From owner-fluorpro@hgmp.mrc.ac.uk Mon May 1 11:41:57 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 29AF317A5C; Mon, 1 May 2000 11:41:54 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id LAA27343 for ; Mon, 1 May 2000 11:41:53 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id LAA15976 for fluorpro-list@hgmp.mrc.ac.uk; Mon, 1 May 2000 11:41:51 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: buubio31@apjapan.co.jp (sam) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Important information for all businesses Date: 1 May 2000 11:41:50 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 29 Message-ID: <200005012810UAA45235@jukmnhgb8j.mil.za> X-Trace: niobium.hgmp.mrc.ac.uk 957177711 15973 193.62.192.80 (1 May 2000 10:41:51 GMT) X-Complaints-To: news@net.bio.net X-Received: from fe00003.mil.za (fe00003.armscor.co.za [196.4.146.103] (may be forged)) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id LAA27317 for ; Mon, 1 May 2000 11:41:46 +0100 (BST) X-Received: from 8gytfg7 ([63.25.15.190]) by fe00003.mil.za (Post.Office MTA v3.5.3 release 223 ID# 0-66616U100L100S0V35) with SMTP id za; Mon, 1 May 2000 06:58:43 +0100 X-To: buubio31@apjapan.co.jp X-Comments: Authenticated sender is To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Are you in need of a Merchant Bank Processing? Don't loose valuable sales by not providing your Customers with the ability to pay by Credit Card! Increase your sales by great numbers $$$ EZ CREDIT Approval Low Transaction Fees Minimum Start-Up Costs On-Line real Time Processing Equipment Lease-Option Plans Deposits Direct To Your Existing Bank Account CardTech Serving the Merchant Banking Industry since 1992 Call Today Toll Free @ 1 (888) 363-7881 and ask for extention #909 For Your FREE consultation! ====================== rem - leave890@yahoo.com ====================== 822680827226942174742 GTo: buubio31@a --- From owner-fluorpro@hgmp.mrc.ac.uk Mon May 1 19:25:51 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id DBDD517A43; Mon, 1 May 2000 19:25:49 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id TAA03345 for ; Mon, 1 May 2000 19:25:47 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA21613 for fluorpro-list@hgmp.mrc.ac.uk; Mon, 1 May 2000 19:25:46 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: diamond@its.caaltech.edu (Rochelle Diamond) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Steven R. Kain Date: 1 May 2000 19:25:45 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 15 Message-ID: <390DCBCD.E86C8991@its.caltech.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 957205546 21611 193.62.192.80 (1 May 2000 18:25:46 GMT) X-Complaints-To: news@net.bio.net X-Received: from chamber.cco.caltech.edu (chamber.cco.caltech.edu [131.215.48.55]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id TAA03324 for ; Mon, 1 May 2000 19:25:38 +0100 (BST) X-Received: from its.caltech.edu (DHCP-35-158.caltech.edu [131.215.35.158]) by chamber.cco.caltech.edu (8.9.3/8.9.3) with ESMTP id LAA17159 for ; Mon, 1 May 2000 11:23:07 -0700 (PDT) X-Mailer: Mozilla 4.7 [en] (Win98; U) X-Accept-Language: en,zh-CN X-To: fluorpro@hgmp.mrc.ac.uk To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk We are trying to reach Steven R. Kain who helped to start this newsgroup. If he is monitoring this list or if anyone knows his whereabouts we would greatly appreciate the help. He contributed a chapter on GFP to our new book from Springer Verlag entitled In Living Color: Protocols in Flow Cytometry and Cell Sorting which was released in March 2000. We are trying to contact him to let him know that we have his complimentary book. He has disappeared off our radar screen. Thanks, Rochelle Diamond, editor diamond@its.caltech.edu 626-395-4947 http://www.nlivingcolor.com --- From owner-fluorpro@hgmp.mrc.ac.uk Wed May 3 12:00:52 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id E2A2917A7B; Wed, 3 May 2000 12:00:50 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id MAA22402 for ; Wed, 3 May 2000 12:00:47 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id MAA08329 for fluorpro-list@hgmp.mrc.ac.uk; Wed, 3 May 2000 12:00:45 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: g.coulton@ic.ac.uk (Gary Coulton) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: ICHC 2000 Date: 3 May 2000 12:00:44 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 76 Message-ID: <3.0.3.32.20000503120034.012b87c0@icex4.cc.ic.ac.uk> Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" X-Trace: niobium.hgmp.mrc.ac.uk 957351645 8326 193.62.192.80 (3 May 2000 11:00:45 GMT) X-Complaints-To: news@net.bio.net X-Received: from judy.ic.ac.uk (judy.ic.ac.uk [155.198.5.28]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id MAA22354 for ; Wed, 3 May 2000 12:00:39 +0100 (BST) X-Received: from juliet.ic.ac.uk ([155.198.5.4]) by judy.ic.ac.uk with esmtp (Exim 2.12 #1) id 12mwtR-0006zG-00 for fluorpro@hgmp.mrc.ac.uk; Wed, 3 May 2000 12:00:37 +0100 X-Received: from icex4.cc.ic.ac.uk ([155.198.3.4]) by juliet.ic.ac.uk with esmtp (Exim 2.12 #1) id 12mwtR-0002aC-00 for fluorpro@hgmp.mrc.ac.uk; Wed, 3 May 2000 12:00:37 +0100 X-Received: from bm-grc1 (bm-grc1.sk.med.ic.ac.uk [155.198.18.63]) by icex4.cc.ic.ac.uk with SMTP (Microsoft Exchange Internet Mail Service Version 5.5.2650.21) id GXD3RAS4; Wed, 3 May 2000 12:00:53 +0100 X-Sender: IC\grc\g.coulton@icex4.cc.ic.ac.uk X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.3 (32) X-To: fluorpro@hgmp.mrc.ac.uk To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Hi All, 11th International Congress of histochemistry and Cytochemistry (ICHC 2000) "Cell Biology Tools for the New Century" ICHC 2000 is the premier meeting this year to address the very latest developments in visualisation techniques for Cell Biology. to be held in the magical medieval city of York, England between 3-8 September 2000. We have a fabulous speaker listof over 100 including some of the biggest names in Cell Biology and Imaging such as Roger Tsien, Richard Haugland, Fred Bosman, Alan Boyde, Stefan Hell, Alan Fine, Jim Smith, Margaret Buckingham, Nick White, Angus Lamond, David Becker, Jeniffer Lipincott-Schwartz and many, many more. Topics include -Tracking Molecules in Cells, Fluorescent markers of Gene Expression, Live Cell Imaging in plants and Fungi, Apoptosis, Cell Signalling, Environmental toxicology, Imaging Embryonic Development, Ion Imaging in Cells, Imaging in the Neurosciences etc., etc. etc. See the science and have a chance to talk to the best in the field. Come for a holiday as well as the science we can offer many great excursions. For more information and details of how to register please visit our web-site at www.med.ic.ac.uk/external/ichc_2000 DON'T MISS OUT IT WILL BE ONE OF THE CONFERENCES OF THE YEAR Dr. Gary Coulton Molecular Pathology Division of Biomedical Sciences Imperial College School of Medicine The Sir Alexander Fleming Building South Kensington London SW7 2AZ tel 0044 (0)171 594 3190 fax 0044 (0)171 594 3022 PLEASE NOTE AS FROM APRIL 1ST 2000 THE FOLLOWING NUMBERS WILL REPLACE THOSE ABOVE AND SHOULD BE USED. TEL. ++44 020 7594 3190 FAX. ++44 020 7594 3022 e-mail g.coulton@ic.ac.uk ------------------------------------- Announcing the 11th International Congress of Histochemistry and Cytochemistry (ICHC 2000) "Understanding Biocomplexity: The Post-Genome Challenge" September 3-8, 2000, York, United Kingdom ICHC 2000 will comprise 27 symposia addressing the latest developments and applications of histochemistry and cytochemistry in the life sciences including medicine. Many leading experts to speak On-line registration now open!!!!!!!!!! For further details of the meeting and how to pre-register please visit our web-site at http://www.med.ic.ac.uk/external/ichc_2000 Hope to see you there. --- From owner-fluorpro@hgmp.mrc.ac.uk Sat May 6 08:40:16 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5451717A6D; Sat, 6 May 2000 08:40:15 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id IAA10763 for ; Sat, 6 May 2000 08:40:13 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id IAA23903 for fluorpro-list@hgmp.mrc.ac.uk; Sat, 6 May 2000 08:40:11 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: freedom4ever@china.com X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: You NEED This Info Date: 6 May 2000 08:40:10 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 102 Message-ID: <200005060736.AAA14137@quartz.airtouch.com> Mime-Version: 1.0 Content-Type: multipart/mixed; boundary="----=_NextPart_000_49A8_00003437.00001EE1" X-Trace: niobium.hgmp.mrc.ac.uk 957598811 23900 193.62.192.80 (6 May 2000 07:40:11 GMT) X-Complaints-To: news@net.bio.net X-Received: from quartz.airtouch.com (quartz.airtouch.com [151.144.253.22]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id IAA10744 for ; Sat, 6 May 2000 08:40:08 +0100 (BST) X-Received: from erik (max1-21.atlanta.corecomm.net [216.214.97.149]) by quartz.airtouch.com (8.8.8+Sun/8.8.8) with SMTP id AAA14137; Sat, 6 May 2000 00:36:34 -0700 (PDT) X-Priority: 3 X-MSMail-Priority: Normal X-Reply-To: nancy@china.com To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk ------=_NextPart_000_49A8_00003437.00001EE1 Content-Type: text/html;
Dear Friend,

Let me ask you a very straightforward question and
before you answer, think about it for a moment.

When you have trouble making ends meet on today's
income, how do you expect to retire and live on
less money than you earn now?

Read the question again before you go on -
it is a very serious question.

If your income stopped today, how many
months could you survive in your current lifestyle
with no money coming in? What will be different in
your life two years from now if you continue
doing what you are doing now?

Corporate America has conditioned us to believe
that security comes from employment. Yet we hope
we never become the victims of downsizing, career
burn out, illness or injury. The frightening
reality for most people is the plastic in their
wallets determines the quality of their lives.

Ask yourself, are you.....

* Tired of working for someone else and getting
paid what THEY tell you you're worth?
* Do your credit card balances total more than
your bank accounts?
* Tired of the get rich quick on $5 fantasy
programs?
* Tired of the Multi Level marketing residual
income illusion?
* Looking for REAL wealth?

CALL NOW 1-800-320-9895 ext 5104
THEN LOOK AT THIS!!!

* Free enterprise in its purest form, not Multi
Level marketing or franchise.
* Proven lead generation system brings qualified
prospects to you.
* Multiple 6 figure lifestyle realistically
attainable in the first year.
* Endless support in an environment of
REAL integrity.
* 80% profit on all sales paid directly to you.

Our team of professionals is looking for a few
quality individuals with a strong work ethic and
DESIRE to generate $2,000 to $5,000 per week from
the comfort of their home. With 5 different income
streams available we teach people how to achieve a
6 figure cash flow within the first 6 to 12 months
and how to retire for good within 2 to 4 years.

SERIOUS SELF-STARTERS WITH A STRONG DESIRE TO IMPROVE
THEIR IMMEDIATE FINANCIAL FUTURE CALL NOW...

CALL NOW 1-800-320-9895 ext 5104

Success is a matter of choice....your choice always.

Prosperous regards!

To be removed from all future mailings reply to
this email Please type remove in the subject line.
Calling the 800 number will not get you removed.
Under Bill s.1618 TITLE III passed by the 105th
U.S. Congress this letter can not be considered
spam as long as we include:
Contact information & a Remove Link

--- From owner-fluorpro@hgmp.mrc.ac.uk Sat May 6 17:27:03 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3994617A7C; Sat, 6 May 2000 17:27:02 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id RAA05351 for ; Sat, 6 May 2000 17:27:00 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA00181 for fluorpro-list@hgmp.mrc.ac.uk; Sat, 6 May 2000 17:26:58 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: urbane45@netscape.net (JJ) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Enzyme kinetics Date: 6 May 2000 17:26:57 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 17 Message-ID: <391456BA.BDE514AC@netscape.net> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 957630418 178 193.62.192.80 (6 May 2000 16:26:58 GMT) X-Complaints-To: news@net.bio.net X-Received: from CDF_NT.college-de-france.fr (mailhub.college-de-france.fr [193.52.22.2]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id RAA05343 for ; Sat, 6 May 2000 17:26:56 +0100 (BST) X-Received: from netscape.net (192.168.7.3 [192.168.7.3]) by CDF_NT.college-de-france.fr with SMTP (Microsoft Exchange Internet Mail Service Version 5.5.2448.0) id KJGV1RH6; Sat, 6 May 2000 18:22:07 +0200 X-Reply-To: urbane45@netscape.net X-Mailer: Mozilla 4.7 (Macintosh; U; PPC) X-To: fluorpro@hgmp.mrc.ac.uk To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Hello, I would like to determine the kinetics of an enzyme in living cells using mutant GFPs as the reporter. I would like to monitor the catalytic activity of the enzyme. I am very much interested in being able to extract quantitative data about the enzymes activity. Does anyone know of any research along these lines using GFPs? Any help would be most helpful. Thank you, JJ urbane45@netscape.net --- From owner-fluorpro@hgmp.mrc.ac.uk Sat May 6 20:52:33 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id F209817A56; Sat, 6 May 2000 20:52:31 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id UAA14082 for ; Sat, 6 May 2000 20:52:30 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id UAA02522 for fluorpro-list@hgmp.mrc.ac.uk; Sat, 6 May 2000 20:52:28 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: urbane45@netscape.net (JJ) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Enzyme kinetics using GFPs Date: 6 May 2000 20:52:27 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 17 Message-ID: <391486E4.73A1BFD5@netscape.net> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 957642748 2519 193.62.192.80 (6 May 2000 19:52:28 GMT) X-Complaints-To: news@net.bio.net X-Received: from CDF_NT.college-de-france.fr (mailhub.college-de-france.fr [193.52.22.2]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id UAA14074 for ; Sat, 6 May 2000 20:52:25 +0100 (BST) X-Received: from netscape.net (192.168.7.3 [192.168.7.3]) by CDF_NT.college-de-france.fr with SMTP (Microsoft Exchange Internet Mail Service Version 5.5.2448.0) id KJGV1R24; Sat, 6 May 2000 21:47:37 +0200 X-Reply-To: urbane45@netscape.net X-Mailer: Mozilla 4.7 (Macintosh; U; PPC) X-To: fluorpro@hgmp.mrc.ac.uk To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Hello, I would like to determine the kinetics of an enzyme in living cells using mutant GFPs as the reporter. I would like to monitor the catalytic activity of the enzyme. I am very much interested in being able to extract quantitative data about the enzymes activity. Does anyone know of any research along these lines using GFPs? Any help would be most helpful. Thank you, JJ urbane45@netscape.net --- From owner-fluorpro@hgmp.mrc.ac.uk Sat May 6 22:12:32 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 7B29E17A56; Sat, 6 May 2000 22:12:31 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id WAA26304 for ; Sat, 6 May 2000 22:12:29 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA12066 for fluorpro-list@hgmp.mrc.ac.uk; Sat, 6 May 2000 22:12:28 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: simon@rockvax.rockefeller.edu (Sanford Simon) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Fwd: Enzyme kinetics Date: 6 May 2000 22:12:26 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 99 Message-ID: <200005062112.RAA15654@rockvax.rockefeller.edu> Mime-Version: 1.0 Content-Type: multipart/alternative; types="text/plain,text/html"; boundary="=====================_24342689==_.ALT" X-Trace: niobium.hgmp.mrc.ac.uk 957647547 12064 193.62.192.80 (6 May 2000 21:12:27 GMT) X-Complaints-To: news@net.bio.net X-Received: from rockvax.rockefeller.edu (rockvax.rockefeller.edu [129.85.1.21]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id WAA26297 for ; Sat, 6 May 2000 22:12:24 +0100 (BST) X-Received: from simon (ppp10.rockefeller.edu [129.85.1.81]) by rockvax.rockefeller.edu (8.9.1/8.9.1) with SMTP id RAA15654 for ; Sat, 6 May 2000 17:12:21 -0400 (EDT) X-Sender: simon@rockvax.rockefeller.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 4.0.2 X-To: fluorpro@hgmp.mrc.ac.uk To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk --=====================_24342689==_.ALT Content-Type: text/plain; charset="us-ascii" Dear JJ, We have recently published a paper doing this: Yu Chen and Sanford M. Simon In Situ Biochemical Demonstration that P-Glycoprotein Is a Drug Efflux Pump with Broad Specificity J. Cell Biol. 2000 148: 863-870. If you have any questions, please feel free to contact us. - Sandy >Subject: Enzyme kinetics >Date: 6 May 2000 17:26:57 +0100 >Hello, > >I would like to determine the kinetics of an enzyme in living cells >using mutant GFPs as the reporter. I would like to monitor the catalytic >activity of the enzyme. I am very much interested in being able to >extract quantitative data about the enzymes activity. > >Does anyone know of any research along these lines using GFPs? > >Any help would be most helpful. > >Thank you, > >JJ >urbane45@netscape.net > >--- > Sandy Simon Laboratory of Cellular Biophysics Box 304 Rockefeller University 1230 York Avenue New York, N.Y. 10021 (212) 327-8130 (voice) (212) 327-8022 (fax) simon@rockvax.rockefeller.edu (e-mail) --=====================_24342689==_.ALT Content-Type: text/html; charset="us-ascii" Dear JJ,

We have recently published a paper doing this:

Yu Chen and Sanford M. Simon
In Situ Biochemical Demonstration that P-Glycoprotein Is a Drug Efflux Pump with Broad Specificity
J. Cell Biol. 2000 148: 863-870.

If you have any questions, please feel free to contact us.

- Sandy

>Subject: Enzyme kinetics
>Date: 6 May 2000 17:26:57 +0100
>Hello,
>
>I would like to determine the kinetics of an enzyme in living cells
>using mutant GFPs as the reporter. I would like to monitor the catalytic
>activity of the enzyme. I am very much interested in being able to
>extract quantitative data about the enzymes activity.
>
>Does anyone know of any research along these lines using GFPs?
>
>Any help would be most helpful.
>
>Thank you,
>
>JJ
>urbane45@netscape.net
>
>---
>

Sandy Simon

Laboratory of Cellular Biophysics

Box 304

Rockefeller University

1230 York Avenue

New York, N.Y. 10021

(212) 327-8130 (voice)

(212) 327-8022 (fax)

simon@rockvax.rockefeller.edu (e-mail) --=====================_24342689==_.ALT-- --- From owner-fluorpro@hgmp.mrc.ac.uk Mon May 8 21:22:44 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 4362317A8B; Mon, 8 May 2000 21:22:43 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id VAA05839 for ; Mon, 8 May 2000 21:22:41 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id VAA02458 for fluorpro-list@hgmp.mrc.ac.uk; Mon, 8 May 2000 21:22:40 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: elee@ncifcrf.gov (mgl-person) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: expression of eGFPcre fusion protein in neuron Date: 8 May 2000 21:22:39 +0100 Organization: ABL Lines: 10 Message-ID: <39171571.56E0B4FE@mail.ncifcrf.gov> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 957817359 2456 193.62.192.80 (8 May 2000 20:22:39 GMT) X-Complaints-To: news@net.bio.net X-Received: from mail.ncifcrf.gov (mail.NCIFCRF.GOV [129.43.100.100]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id VAA05828 for ; Mon, 8 May 2000 21:22:36 +0100 (BST) X-Received: from mail.ncifcrf.gov ([129.43.18.95]) by mail.ncifcrf.gov (8.10.1/8.10.1) with ESMTP id e48KNEL26157 for ; Mon, 8 May 2000 16:23:15 -0400 (EDT) X-Mailer: Mozilla 4.6 (Macintosh; U; PPC) X-Accept-Language: en X-To: fluorpro@hgmp.mrc.ac.uk To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk I made transgenic mice carrying neuron-specific promoter-driven eGFPcre fusion protein. Cre functions very well. I detected in most of adult neurons. However, I couldn't detect eGFP signal although I checked this contruct in in vitro culture cells and it showed very strong eGFP signal. I used 4% cold paraformaldehyde to do perfusion and the other 3-hr fixation in the same fixative. I think eGFP won't lose in this way. I wonder if anybody has an experience in expressing eGFP in adult neurons of transgenic mice. Please give some suggestion. --- From owner-fluorpro@hgmp.mrc.ac.uk Wed May 10 13:48:10 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 1747D17A4E; Wed, 10 May 2000 13:48:08 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id NAA02424 for ; Wed, 10 May 2000 13:48:07 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA10812 for fluorpro-list@hgmp.mrc.ac.uk; Wed, 10 May 2000 13:48:06 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Piers Mahon X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: DUAL CFP/YFP RATIO IMAGING Date: Wed, 10 May 2000 13:54:32 +0100 Organization: University of Cambridge, England Lines: 21 Message-ID: <39195C08.959DCB70@mole.bio.cam.ac.uk> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.03 [en] (WinNT; I) To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk I am about to try quantitative ratio imaging on a TCS using CYAN and YELLOW fluorescent proteins with different sub-cellular location signals. I am expecting any positive results to be very subtle images differences (i.e. not blue and yellow blobs in different parts of the cell, but a cyan/yellow signal ratio that varies in sub-sections of the same blob). If you have any experience of CYAN FP/ YELLOW FP imaging, or related dual imaging, please get in touch. I'm particularly interested in advice in three areas: 1) selecting the right filter ranges to obtain the best bleed through / sensitivity compromise, 2) Indications of the random noise levels I can expect in my negative controls of dual transformants containing CYAN and YELLOW with the same location signals, 3) Suggestions for statistical tests that I can perform to see if ratio images with different signals on the two reporters are more variable than images captured using the same signal. Many thanks in advance, Piers Mahon From owner-fluorpro@hgmp.mrc.ac.uk Thu May 11 06:08:37 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C971E17A7F; Thu, 11 May 2000 06:08:35 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id GAA25148 for ; Thu, 11 May 2000 06:08:34 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id GAA01611 for fluorpro-list@hgmp.mrc.ac.uk; Thu, 11 May 2000 06:08:33 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: keagea38@kurasc.kyoto-u.ac.jp (Mary) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Go Anywhere ! Date: 11 May 2000 06:08:32 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 40 Message-ID: <200005113522PAA10706@lkijuhyg6a.fra.fr> X-Trace: niobium.hgmp.mrc.ac.uk 958021713 1609 193.62.192.80 (11 May 2000 05:08:33 GMT) X-Complaints-To: news@net.bio.net X-Received: from saturne (ns.fra.fr [194.2.70.1]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with SMTP id GAA25141 for ; Thu, 11 May 2000 06:08:30 +0100 (BST) X-Received: from sdn-ar-002papittP130.dialsprint.net (sdn-ar-002papittP130.dialsprint.net [168.191.211.194]) by saturne.fra.fr (NTMail 3.03.0013/4c.abax) with ESMTP id sa130096 for ; Thu, 11 May 2000 06:28:51 +0200 X-To: keagea38@kurasc.kyoto-u.ac.jp X-Comments: Authenticated sender is To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk INTERNATIONAL DRIVER'S LICENSE Need a new driver's license? Too many points or other trouble? Want a license that can never be suspended or revoked? Want ID for nightclubs or hotel check-in? Avoid tickets, fines, and mandatory driver's education. Protect your privacy, and hide your identity. The United Nations gave you the privilege to drive freely throughout the world! (Convention on International Road Traffic of September 19, 1949 & World Court Decision, The Hague, Netherlands, January 21, 1958) Take advantage of your rights. Order a valid International Driver's License that can never be suspended or revoked. Confidentiality assured. CALL NOW!!! 1-937-586-9313 rem- detroit89@earthdome.com 210950821003169430108402 u[ Nethe --- From owner-fluorpro@hgmp.mrc.ac.uk Thu May 11 16:02:11 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 1EB7117A43; Thu, 11 May 2000 16:02:06 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id QAA22079 for ; Thu, 11 May 2000 16:02:04 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA09321 for fluorpro-list@hgmp.mrc.ac.uk; Thu, 11 May 2000 16:02:03 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Jerry X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Re: DUAL CFP/YFP RATIO IMAGING Date: Thu, 11 May 2000 09:46:52 -0400 Organization: The University of North Carolina at Chapel Hill Lines: 45 Message-ID: <391AB9CC.234DB2A8@med.unc.edu> References: <39195C08.959DCB70@mole.bio.cam.ac.uk> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: news2.isis.unc.edu 958053056 7278 152.19.37.137 (11 May 2000 13:50:56 GMT) X-Complaints-To: usenet@news2.isis.unc.edu X-Mailer: Mozilla 4.61 [en] (Win98; I) X-Accept-Language: en To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Hello, I chose filters for emerald/topaz imaging. To choose the excitation filters I took ratio of the relative excitation of emerald and topaz at each wavelength. The minimum ratio occurs at the optimum wavelength for exciting one fluorophore and the maximum ratio occurs at the optimum wavelength for exciting the other. The same can be done for the emission filters. This is only a starting point since the signal size will be small with very narrow band filters. I incorporated wavelengths moving away from the optimum. The ratio to use for multiple wavelengths is the ratio of the sums of the relative excitations (or emissions) (the ratio of the areas under the curves). At some point the limit on the filter pass bands may be that the excitation and emission filter must have enough space between them to accomodate the dichroic mirror. If you desire your measurements to be maximally precise you might need to consider the possibilty of FRET between cyan and yellow FP. The article by Gordon and Herman in Biophysical Journal May 1998 gives methods for estimating the concentrations of two fluorophores in the presence of FRET and crosstalk. (calculation of FRET requires estimates of the fluorophore concentrations or the ratio of concentrations) HTH, Jerry gordon@med.unc.edu Piers Mahon wrote: > > I am about to try quantitative ratio imaging on a TCS using CYAN and > YELLOW fluorescent proteins with different sub-cellular location > signals. I am expecting any positive results to be very subtle images > differences (i.e. not blue and yellow blobs in different parts of the > cell, but a cyan/yellow signal ratio that varies in sub-sections of the > same blob). > > If you have any experience of CYAN FP/ YELLOW FP imaging, or related > dual imaging, please get in touch. I'm particularly interested in > advice in three areas: 1) selecting the right filter ranges to obtain > the best bleed through / sensitivity compromise, 2) Indications of the > random noise levels I can expect in my negative controls of dual > transformants containing CYAN and YELLOW with the same location signals, > 3) Suggestions for statistical tests that I can perform to see if ratio > images with different signals on the two reporters are more variable > than images captured using the same signal. > > Many thanks in advance, > > Piers Mahon From owner-fluorpro@hgmp.mrc.ac.uk Wed May 17 11:10:28 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C76E817A63; Wed, 17 May 2000 11:10:26 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id LAA15711 for ; Wed, 17 May 2000 11:10:24 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id LAA10702 for fluorpro-list@hgmp.mrc.ac.uk; Wed, 17 May 2000 11:10:22 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f Message-ID: <392270DC.28E80BC3@nowhere.com> Date: Wed, 17 May 2000 12:13:53 +0200 From: Hansi Reply-To: hase@nowhere.com X-Mailer: Mozilla 4.61 (Macintosh; I; PPC) X-Accept-Language: en MIME-Version: 1.0 X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Re: ds RED References: <8djqp9$1lra$1@www.univie.ac.at> Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit X-Trace: 17 May 2000 12:10:15 +0100, berlepsch.ethz.ch Organization: Swiss Federal Institute of Technology (ETHZ) Lines: 45 To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk We have used dsRed both on its own and fused to other proteins and found that the intensity is equal to or greater than that of GFP. Also, there is no obvious difference in expression levels. We haven't tried to produce any stable lines yet. Daniel Martin Offterdinger wrote: > Hi, > Is there anyone who has already used dsRED as an alternative to EGFP. > What are your experiences in terms of > -fluorescence intensity > -generation of stable cell lines > -expression levels > compared to EGFP-fusions? > Any comments are highly appreciated > martin -- ((((((((((((((((((((((((((((())))))))))))))))))))))))))))) Daniel Auerbach Institute Of Cell Biology ETH Hoenggerberg 8093 Zuerich, Switzerland Phone++41 633 33 93 Fax++41 633 10 69 auerbach@cell.biol.ethz.ch Visit our group at: http://www.cell.biol.ethz.ch/structure/perriard/research.html There was a beer makers convention going on, and after the days meetings several beer company presidents decided to go to a bar for some drinks. So, the Budweiser president orders a Bud, the Coors president orders a Coors, The Miller president orders a Millers and so on. When it comes to the Guinness President, he looks at the group of men and then orders a glass of water. The rest of the presidents look at him and ask "Why aren't you ordering a Guinness?" and he looked at them and replied, "Lads, if you're not going to drink beer then I'm not going to either." ((((((((((((((((((((((((((((()))))))))))))))))))))))))))))) From owner-fluorpro@hgmp.mrc.ac.uk Wed May 17 14:56:14 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 05F6117A59; Wed, 17 May 2000 14:56:12 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id OAA08409 for ; Wed, 17 May 2000 14:56:11 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA13572 for fluorpro-list@hgmp.mrc.ac.uk; Wed, 17 May 2000 14:56:09 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: bachelorcd@uswest.net X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: For Serious Marketers ONLY! Date: 17 May 2000 14:56:07 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 40 Message-ID: <200005171347.WAA20017@whict.edu.cn> X-Trace: niobium.hgmp.mrc.ac.uk 958571768 13570 193.62.192.80 (17 May 2000 13:56:08 GMT) X-Complaints-To: news@net.bio.net X-Received: from whict.edu.cn ([210.42.24.20]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with SMTP id OAA08384 for ; Wed, 17 May 2000 14:55:59 +0100 (BST) X-Received: from lister by whict.edu.cn (SMI-8.6/SMI-SVR4) id WAA20017; Wed, 17 May 2000 22:47:07 +0900 X-To: jonsell@dunlopdriver.com To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk For Serious Marketers ONLY! Direct Email Marketing Are you making more money that ever? Are You getting all the sales that you want? Is your inbox filled with fresh orders every day? If you answered YES to all of the above questions, then you probably don't need our services. However, If you answered No to ANY of them, then try Warhorse Promotionz! Warhorse.... IS YOUR COMPLETE ANSWER! We are Contract Mailers with Quality Lists, that have been cleaned and verified. These lists get 90% plus deliverables. Contract Mailing We can send your ad to 100,000 or to 10 million We capture all replies, including removes and flames We send you only the qualified leads! Bulk Friendly Hosting We can host a form site to enable you to capture and retrieve all your leads *NO TARGETED LISTS * NO TARGETED LISTS * *NO TARGETED LISTS * NO TARGETED LISTS * To find out how to get the leads that you NEED, Call 953 567-8552, Leave your name, phone # and Clearly spell out your email address We WILL return your call ASAP --- From owner-fluorpro@hgmp.mrc.ac.uk Thu May 18 07:14:07 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5635417AD0; Thu, 18 May 2000 07:14:06 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id HAA03740 for ; Thu, 18 May 2000 07:14:04 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id HAA05259 for fluorpro-list@hgmp.mrc.ac.uk; Thu, 18 May 2000 07:14:02 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: k.sunn@garvan.unsw.edu.au (Kathryn Sunn) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: GFP bleed thru in double labelling Date: 18 May 2000 07:14:01 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 31 Message-ID: Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" X-Trace: niobium.hgmp.mrc.ac.uk 958630441 5256 193.62.192.80 (18 May 2000 06:14:01 GMT) X-Complaints-To: news@net.bio.net X-Received: from gimr.garvan.unsw.edu.au (gimr.garvan.unsw.EDU.AU [129.94.136.101]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id HAA03732 for ; Thu, 18 May 2000 07:13:55 +0100 (BST) X-Received: from [129.94.42.138] ([129.94.42.138]) by gimr.garvan.unsw.edu.au (8.9.1/8.9.1) with ESMTP id QAA05254 for ; Thu, 18 May 2000 16:17:37 +1000 (EST) X-Sender: katsun@gimr.garvan.unsw.edu.au X-To: fluorpro@hgmp.mrc.ac.uk To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Dear All I was wondering if anyone has had any experience with bleed through of fluorescence from the EGFP channel, when looking at an antibody on the red channel? I have transfected cells with the EGFP fusion constructs and then wish to look at colocalision with another protein by immunocytochemistry. I am using an ALEXA-594 secondary antibody, but I cant seem to detect an intense specific red signal as the green fluorecence is bleeding through to the red on the confocal. Has anyone had this happen to them, how did you overcome this problem? I am trying to fix (in 2%PFA) at 4 degrees, and also was with 0.1% sodium borohydride to see if this helps, but dont know the results as yet. If anyone could help I would greatly appreciate it. Thanks Kate ___________________________________________________________________ Kathryn Sunn BSc (Hons) Bone and Mineral Research Program Garvan Institute of Medical Research St Vincent's Hospital Tel: (02) 295 8260 Sydney NSW 2010 Fax: (02) 295 8241 Australia email: k.sunn@garvan.unsw.edu.au ___________________________________________________________________ --- From owner-fluorpro@hgmp.mrc.ac.uk Thu May 18 10:25:09 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id B974D17A6F; Thu, 18 May 2000 10:25:07 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id KAA19760 for ; Thu, 18 May 2000 10:25:05 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id KAA07627 for fluorpro-list@hgmp.mrc.ac.uk; Thu, 18 May 2000 10:25:04 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Cornelius Krasel Subject: Re: GFP bleed thru in double labelling X-Newsgroups: bionet.molbio.proteins.fluorescent References: Organization: =?ISO-8859-1?Q?Universit=E4t_W=FCrzburg?= User-Agent: tin/pre-1.4-19990805 ("Preacher Man") (UNIX) (Linux/2.2.13 (i686)) Date: Thu, 18 May 2000 11:16:43 +0200 Message-ID: X-Trace: 18 May 2000 11:25:01 +0100, wpxx02.toxi.uni-wuerzburg.de Lines: 21 To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Kathryn Sunn wrote: > I was wondering if anyone has had any experience with bleed through of > fluorescence from the EGFP channel, when looking at an antibody on the red > channel? I have transfected cells with the EGFP fusion constructs and then > wish to look at colocalision with another protein by immunocytochemistry. I > am using an ALEXA-594 secondary antibody, but I cant seem to detect an > intense specific red signal as the green fluorecence is bleeding through to > the red on the confocal. Has anyone had this happen to them, how did you > overcome this problem? I cannot confirm that. I used various (green) GFPs and could successfully colocalize them with Alexa-594 without any bleedthrough. I fix my cells in PBS / 5% Sucrose / 3% PFA at room temperature for 20 minutes and mount them in 60% glycerol in PBS without any antifade agent. --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP4 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-fluorpro@hgmp.mrc.ac.uk Thu May 18 14:33:10 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 4870917A88; Thu, 18 May 2000 14:33:08 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id OAA17698 for ; Thu, 18 May 2000 14:33:05 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA10785 for fluorpro-list@hgmp.mrc.ac.uk; Thu, 18 May 2000 14:33:03 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: bludin@lis.ch (Beat Ludin) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Re: GFP bleed thru in double labelling Date: 18 May 2000 14:33:02 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 48 Message-ID: References: Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" ; format="flowed" X-Trace: niobium.hgmp.mrc.ac.uk 958656782 10782 193.62.192.80 (18 May 2000 13:33:02 GMT) X-Complaints-To: news@net.bio.net X-Received: from gamma.dynaweb.de ([195.127.244.2]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id OAA17681 for ; Thu, 18 May 2000 14:32:53 +0100 (BST) X-Received: from [195.24.89.153] ([195.24.89.153]) by gamma.dynaweb.de (8.9.1/8.9.1) with ESMTP id PAA03113; Thu, 18 May 2000 15:32:29 +0200 (CEST) X-Sender: lis3@pop.lis.ch X-In-Reply-To: X-To: k.sunn@garvan.unsw.edu.au (Kathryn Sunn) X-Cc: GFP Mailing Llist To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk What filters/settings are you using? Do you record the two channels simultaneously (emission filtering only) or sequentially (excitation and emission filtering)? Beat >Dear All > >I was wondering if anyone has had any experience with bleed through of >fluorescence from the EGFP channel, when looking at an antibody on the red >channel? I have transfected cells with the EGFP fusion constructs and then >wish to look at colocalision with another protein by immunocytochemistry. I >am using an ALEXA-594 secondary antibody, but I cant seem to detect an >intense specific red signal as the green fluorecence is bleeding through to >the red on the confocal. Has anyone had this happen to them, how did you >overcome this problem? > >I am trying to fix (in 2%PFA) at 4 degrees, and also was with 0.1% sodium >borohydride to see if this helps, but dont know the results as yet. > >If anyone could help I would greatly appreciate it. > >Thanks >Kate > >___________________________________________________________________ >Kathryn Sunn BSc (Hons) >Bone and Mineral Research Program >Garvan Institute of Medical Research >St Vincent's Hospital Tel: (02) 295 8260 >Sydney NSW 2010 Fax: (02) 295 8241 >Australia email: k.sunn@garvan.unsw.edu.au >___________________________________________________________________ > > > >--- LIFE IMAGING SERVICES - visit our web site at http://www.lis.ch +----------------------------------------------------------- | Dr. B. Ludin | Life Imaging Services fon ++41 (0)79 235 7154 | Muehletalweg 22 fax ++41 (0)860 622 963 160 | CH-4600 Olten beat.ludin@lis.ch | Switzerland http://www.lis.ch --- From owner-fluorpro@hgmp.mrc.ac.uk Thu May 18 15:50:20 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id B967317A88; Thu, 18 May 2000 15:50:18 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id PAA26592 for ; Thu, 18 May 2000 15:50:11 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA11823 for fluorpro-list@hgmp.mrc.ac.uk; Thu, 18 May 2000 15:50:10 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Michael C. Wussow" X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Re: GFP bleed thru in double labelling Date: Thu, 18 May 2000 09:44:44 -0500 Organization: University of Wisconsin, Madison Lines: 57 Message-ID: References:

Reply-To: wussow@bitplane.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: news.doit.wisc.edu 958661025 16856 128.104.136.84 (18 May 2000 14:43:45 GMT) X-Complaints-To: abuse@doit.wisc.edu X-Newsreader: Forte Agent 1.8/32.548 To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk The likely cause is the filters in the confocal. Look at the emission wavelength of both the EGFP and the ALEXA-594 (remember there are a whole bunch of different varieties that emmit at different wavelengths even within the same characterized name so check). www.omegafiters.com or www.molecularprobes.com (good places to check) I have found EGFP excited at from as low as 350 to a max of 525 with a peak at somewhere just under 500nm. The EGFP I have found emmits between 480 all the way up to 600nm with a peak somewhere near 510 nm. The Alexa 594 can be excited way down below 350 nm all the way up to 600 nm with a peak at 590 ish. The Alexa 594 emmits somewhere between 575 and over 700 nm with a peak somewhere near 615 nm. To get rid of the GFP bleed through make sure the emission filter for the Alexa 594 is over 600. You loose some signal but you cut down the bleed through from GFP. The alternative is to move the excitation of the Alexa up to above 525nm. Therefore you do not excite much EGFP and you still get Alexa excitation. Since you mentioned colocalization I encourage you to check out www.bitplane.com we have some nice software for colocalization. Choose products and then colocalization. Good Luck On Thu, 18 May 2000 11:16:43 +0200, Cornelius Krasel wrote: >Kathryn Sunn wrote: >> I was wondering if anyone has had any experience with bleed through of >> fluorescence from the EGFP channel, when looking at an antibody on the red >> channel? I have transfected cells with the EGFP fusion constructs and then >> wish to look at colocalision with another protein by immunocytochemistry. I >> am using an ALEXA-594 secondary antibody, but I cant seem to detect an >> intense specific red signal as the green fluorecence is bleeding through to >> the red on the confocal. Has anyone had this happen to them, how did you >> overcome this problem? > >I cannot confirm that. I used various (green) GFPs and could successfully >colocalize them with Alexa-594 without any bleedthrough. I fix my cells >in PBS / 5% Sucrose / 3% PFA at room temperature for 20 minutes and mount >them in 60% glycerol in PBS without any antifade agent. > >--Cornelius. From owner-fluorpro@hgmp.mrc.ac.uk Thu May 18 16:33:57 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id A283717A88; Thu, 18 May 2000 16:33:52 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id QAA01567 for ; Thu, 18 May 2000 16:33:47 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA12350 for fluorpro-list@hgmp.mrc.ac.uk; Thu, 18 May 2000 16:33:46 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: akohl001@udcf.gla.ac.uk (Alain Kohl) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Proteasome/Renilla Luc. Date: 18 May 2000 16:33:44 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 16 Message-ID: <3.0.6.32.20000518163557.007947a0@udcf.gla.ac.uk> Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" X-Trace: niobium.hgmp.mrc.ac.uk 958664026 12348 193.62.192.80 (18 May 2000 15:33:46 GMT) X-Complaints-To: news@net.bio.net X-Received: from lenzie.cent.gla.ac.uk (lenzie.cent.gla.ac.uk [130.209.16.18]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id QAA01545 for ; Thu, 18 May 2000 16:33:41 +0100 (BST) X-Received: from lab-a421.vir.gla.ac.uk (lab-a421.vir.gla.ac.uk [130.209.134.117]) by lenzie.cent.gla.ac.uk (8.9.3/8.9.3) with SMTP id QAA10573 for ; Thu, 18 May 2000 16:33:39 +0100 (BST) X-Sender: akohl001@udcf.gla.ac.uk (Unverified) X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) X-To: fluorpro@hgmp.mrc.ac.uk To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Dear All, does anybody know if proteasome inhibitors such as MG132 have an effect on renilla or firefly luciferase? We are using those genes as reporters and observe activity decrease even after short in incubations...it's a bit a mistery to us right now, so if anybody knows which secundary effect of MG132 might be responsible, let me know. Thanks Alain Dr Alain Kohl Division of Virology University of Glasgow Church Street Glasgow G3 6LF Scotland, UK --- From owner-fluorpro@hgmp.mrc.ac.uk Sun May 21 22:28:48 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5085E17A61; Sun, 21 May 2000 22:28:47 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id WAA10169 for ; Sun, 21 May 2000 22:28:45 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA12401 for fluorpro-list@hgmp.mrc.ac.uk; Sun, 21 May 2000 22:28:44 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: cablemania@china.com X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: adv: Cable Descrambler Plans Date: 21 May 2000 22:28:43 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 94 Message-ID: <200005212119.FAA62861@smtp4.singnet.com.sg> X-Trace: niobium.hgmp.mrc.ac.uk 958944524 12399 193.62.192.80 (21 May 2000 21:28:44 GMT) X-Complaints-To: news@net.bio.net X-Received: from smtp4.singnet.com.sg (smtp4.singnet.com.sg [165.21.101.224]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id WAA10131 for ; Sun, 21 May 2000 22:28:40 +0100 (BST) X-Received: from china.com (ad202.166.13.39.magix.com.sg [202.166.13.39]) by smtp4.singnet.com.sg (8.9.3/8.9.3) with SMTP id FAA62861; Mon, 22 May 2000 05:19:14 +0800 (SGT) (envelope-from cablemania@china.com) X-To: cablemania@china.com To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk NOTE: THIS IS AN ADVERTISEMENT FOR CABLE TV DESCRAMBLER PLANS IF YOU HAVE NO INTEREST IN THIS INFORMATION PLEASE CLICK DELETE NOW. ARE TO BE REMOVED FROM FUTURE MAILINGS FOLLOW THE DIRECTIONS AT THE END OF MESSAGE moreno68@flashmail.com THANK YOU -========================================================================= CABLE TV DE-SCRAMBLER Want to watch Sporting Events?--Movies?--Pay-Per-View?? *This is the Famous Descrambler Plans You can get the parts for about $12.00 from any electronics store. We Send You: E-Z To follow Assembly Instructions. E-Z TO read Original Drawings. The Parts List. PLUS SOMETHING NEW YOU MUST HAVE! Something you can't do without. THE UP-TO-DATE REPORT:USING A DESCRAMBLER Warning: You should not build a TV Descrambler without reading this report first. Frequently Asked Questions--CABLE TV DESCRAMBLER Q:Will a descrambler work on a Fiber, TCI, Jarrod and satellite system? A:The answer is Yes. In respect to satellite,you just get more stuff! There is one exception:The descrambler will not work with DSS satillite. Q: Do I need a converter box? A: This plan works with or without a converter box. Specific in stuctions are included in the plans for for each! Q: Can the cable company detect that I have the descrambler? A: No,the signal descramblers right at the box and does not move back through the line! Q: Do I have to alter my existing cable system, television or VCR? A: The answer is no! Q: Does this work with my remote control? A: The answer is yes. The descrambler is manually controlled--but very easy to use! Q: Can you email me the plans? A: No the program comes with an easy to follow picture guide. Q: Does this work everywhere across the country? A: Yes,every where in the USA Plus England ,Brazil, Canada and other countries! Q: Is this deal guaranteed? A: Yes if you are unhappy for any reason we will refund your money. ORDER INFORMATION =================================================================== ACT WITHIN THE NEXT 7 DAYS AND RECEIVE TWO FREE BONUSES !!! THE CABLE MANUAL! This manual contains hard to find information your cable company does not want you to know. Also receive The RADAR JAMMER PLANS! Never get another speeding ticket. Build your own radar jammer,this unit will jam police radar so they can't get a reading on your vechicle. Radar jammers are legal in 48 states. It is simple to build.The FREE BONUSES ALONE ARE WORTH ACTING NOW! THE BONUSES FOR FREE. YOU HAVE NOTHING TO LOSE ONLY FREE CABLE TV TO GAIN! ACT NOW! SIMPLY SEND $14.00 CHECK OR, MONEY ORDER , OR CREDIT CARD INFORMATION TO: INFORMATION FACTORY PO BOX 795 NORTH PLATTE NE 69103. For all major credit card orders, please fill out the form bellow and mail in or if you want to be deleted from mail please put REMOVE in subject line so are software can delete you. Thank ACCOUNT NUMBER_________________________ EXP.DATE_______________________________ NAME_______________________ BILLING ADDRESS____________________________ CITY/STATE/ZIP_____________________________ HOME PHONE_________________________________ ****************Remove Instructions******************** ******************************************************** To be removed from future mailings!!!! All REMOVE requests AUTOMATICALLY honored upon receipt. mailto moreno68@flashmail.com subject=REMOVE PLEASE understand that any effort to disrupt, close or block this REMOVE account can only result in difficulties for others wanting to be removed from our mailing list as it will be impossible to take anyone off the list if the remove instruction is not received. ******************************************************* --- From owner-fluorpro@hgmp.mrc.ac.uk Wed May 24 14:17:13 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 31E1A17ADF; Wed, 24 May 2000 14:17:12 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 6937F17ADE; Wed, 24 May 2000 14:17:10 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id C9ED2415D5 for ; Wed, 24 May 2000 14:17:09 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA17141 for fluorpro-list@hgmp.mrc.ac.uk; Wed, 24 May 2000 14:17:09 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: hhtom23@bizland.com ("Henry Johns") X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Your Web #3468 Date: 24 May 2000 14:17:08 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 36 Message-ID: <20000524131705.25C4D415D5@mercury.hgmp.mrc.ac.uk> Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable X-Trace: niobium.hgmp.mrc.ac.uk 959174229 17139 193.62.192.80 (24 May 2000 13:17:09 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id DA7E417ADE; Wed, 24 May 2000 14:17:07 +0100 (BST) X-Received: from server-conam.conam.gob.pe (conam.gob.pe [200.37.42.194]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 25C4D415D5 for ; Wed, 24 May 2000 14:17:05 +0100 (BST) X-Received: from host ([206.133.176.41]) by server-conam.conam.gob.pe (Post.Office MTA v3.0 release 0122 ID# 0-43299U100L2S100) with ESMTP id AAA82; Wed, 24 May 2000 08:18:12 -0500 X-To: out39d@hgmp.mrc.ac.uk X-Mailer: Mozilla 4.70 [en] (Win95; I) To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk WE MAKE IT EASY & A=46=46ORDABLE TO ACCEPT CREDIT CARDS =46OR YOUR BUS= INESS ! INTERNET (Auction Vendors & Online Mall Stores Too!) STORE=46RONT OR MAIL ORDER MERCHANTS WE SPECIALIZE IN APPROVING YOU! APPLY TODAY AND START =46OR JUST $9.95! =46REE APPLICATION!! =46REE PROGRAMMING!! DON'T LOSE ANOTHER SALE! APPLY TO ACCEPT CREDIT CARDS AND CALL (888) 264-9272 DON'T =46ORGET TO ASK ABOUT OUR WEB DESIGN AND HOSTING PACKAGE !!! ************************************************************ If you receive this message and have never joined one of our email lists you can be removed by replying to: mailto:keyst2p@netscape.net?subject=3Dremove ************************************************************ --- From owner-fluorpro@hgmp.mrc.ac.uk Wed May 24 17:00:25 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 27D8217B75; Wed, 24 May 2000 17:00:23 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 4B9EC17B74; Wed, 24 May 2000 17:00:21 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 9B870415CA for ; Wed, 24 May 2000 17:00:20 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA19222 for fluorpro-list@hgmp.mrc.ac.uk; Wed, 24 May 2000 17:00:20 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Jos Broers X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Re: GFP bleed thru in double labelling Date: Wed, 24 May 2000 17:42:21 +0200 Organization: Universiteit Maastricht Lines: 71 Message-ID: <392BF85D.4E7D5B89@molcelb.unimaas.nl> References: Mime-Version: 1.0 Content-Type: multipart/mixed; boundary="------------968008C33CBD9B43597BC8B5" X-Mailer: Mozilla 4.72 (Macintosh; I; PPC) X-Accept-Language: en To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk This is a multi-part message in MIME format. --------------968008C33CBD9B43597BC8B5 Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit Dear Kathryn, If you use the right filter set bleed through of EGF into the red channel should not be a serious problem. We use a rather narrow bandpass filter for GFP (the same one which we use also for FITC) and use TexasRed with a LP filter for the immunolabeling and encounter no serious bleed through unless the GFP signal is extremely bright. Fixation with PFA can cause autofluorescence, but you easily distinguish this from the GFP signal , since you will notice that also in areas of the cell where you don't have GFP signal. Kind regards Jos Kathryn Sunn wrote: > Dear All > > I was wondering if anyone has had any experience with bleed through of > fluorescence from the EGFP channel, when looking at an antibody on the red > channel? I have transfected cells with the EGFP fusion constructs and then > wish to look at colocalision with another protein by immunocytochemistry. I > am using an ALEXA-594 secondary antibody, but I cant seem to detect an > intense specific red signal as the green fluorecence is bleeding through to > the red on the confocal. Has anyone had this happen to them, how did you > overcome this problem? > > I am trying to fix (in 2%PFA) at 4 degrees, and also was with 0.1% sodium > borohydride to see if this helps, but dont know the results as yet. > > If anyone could help I would greatly appreciate it. > > Thanks > Kate > > ___________________________________________________________________ > Kathryn Sunn BSc (Hons) > Bone and Mineral Research Program > Garvan Institute of Medical Research > St Vincent's Hospital Tel: (02) 295 8260 > Sydney NSW 2010 Fax: (02) 295 8241 > Australia email: k.sunn@garvan.unsw.edu.au > ___________________________________________________________________ > > --- --------------968008C33CBD9B43597BC8B5 Content-Type: text/x-vcard; charset=us-ascii; name="Jos.Broers.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Jos Broers Content-Disposition: attachment; filename="Jos.Broers.vcf" begin:vcard n:Broers;Jos tel;work:+31-433881366/1364 x-mozilla-html:FALSE version:2.1 email;internet:Jos.Broers@molcelb.unimaas.nl adr;quoted-printable:;;University Maastricht=0D=0ADept.of Molecular Cell Biology & Genetics=0D=0AP.O. Box 616=0D=0A6200 MD Maastricht=0D=0AThe Netherlands;;;; x-mozilla-cpt:;1 fn:Jos Broers end:vcard --------------968008C33CBD9B43597BC8B5-- From owner-fluorpro@hgmp.mrc.ac.uk Fri May 26 01:12:01 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C9B9B17AC2; Fri, 26 May 2000 01:11:59 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 11C2F17A96; Fri, 26 May 2000 01:11:57 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 509B7415DD for ; Fri, 26 May 2000 01:11:57 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id BAA00543 for fluorpro-list@hgmp.mrc.ac.uk; Fri, 26 May 2000 01:11:56 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: samantha543@mpg-net.de X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Earn $50,000 in just 90 Days! Date: 26 May 2000 01:11:55 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 548 Message-ID: <200005280702.OAA26371@ns.bogor.indo.net.id> Mime-Version: 1.0 Content-Type: text/plain; charset=unknown-8bit X-Trace: niobium.hgmp.mrc.ac.uk 959299916 541 193.62.192.80 (26 May 2000 00:11:56 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 672D017A96; Fri, 26 May 2000 01:11:53 +0100 (BST) X-Received: from ns.bogor.indo.net.id (ns.bogor.indo.net.id [202.159.24.46]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id E6097415DD for ; Fri, 26 May 2000 01:11:46 +0100 (BST) X-Received: from monorailpc (host-216-77-211-157.fll.bellsouth.net [216.77.211.157]) by ns.bogor.indo.net.id (8.8.5/8.8.5) with SMTP id OAA26371; Sun, 28 May 2000 14:02:34 +0700 X-To: samantha543@mpg-net.de To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk So, how in the world have you been? Do you remember holding previous conversations regarding business and money making opportunities. I did not send this to you in error! You said: "If only I could find an easier way to make a higher income" and "If I had more money, I could spend more time with my Family, and less time at work" and "I sure could use more money so I could pay off my bills once and for all!" and "I would love to get involved in a business in which will generate money while I am not at work (like a Gas Pump)" Dear Friend, There is a possibility that we haven't met, but you were chosen by someone to receive this E-Mail. Please, please, print this off and read thoroughly. Be sure that you don't miss any of the points outlined. Then put it down, and then read it again. I am sending you a whole lot of information in which you might not understand the first time you read it. If you don't believe this program will work for you, send it to 10-20 of your closest friends (in which you trust deeply), and ask them what they think? This really works! Have the faith, don't miss this opportunity, get involved also, and it will work for you as it does for us!!!!! Due to the popularity of this letter on the Internet, $$$ A MAJOR NIGHTLY NEWS Program recently dedicated an entire show to the investigation of the program described below to see if it really can make people money. The show also investigated whether or not the program was legal. Their findings proved that there are absolutely no laws prohibiting the participation in the program. This has helped to show people that this is a simple, harmless and fun way to make some extra money at home. The results have been truly remarkable. So many people are participating that those involved are doing much better than ever before. Since everyone makes more as more people try it out, its been very exciting. $$$ You will understand only if you get involved! ********* THE ENTIRE PLAN IS HERE BELOW ********* *** Print This Now For Future Reference *** $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ If you would like to make AT LEAST $50,000 in less than 90 days! If not, forward this to someone who would like to make this kind of money....... It works (like designed) but only for those who follow it to the letter! Please read this program...THEN READ IT AGAIN!! $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ THIS IS A LEGITIMATE, LEGAL, MONEY MAKING OPPORTUNITY!! It does NOT require you to come into contact with people or make or take any telephone calls. Just follow the instructions, and you will make money. This simplified e-mail marketing program works perfectly 100% EVERY TIME! E-mail is the sales tool of the future. Take advantage of this virtually free method of advertising NOW!!! The longer you wait, the more people will be doing business using email. Get your piece of this action!!! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Hello - My name is Johnathon Rourke, I'm from Rhode Island. The enclosed information is something I almost let slip through my fingers. Fortunately, sometime later I re-read everything and gave some thought and study to it. Two years ago, the corporation I worked for the past twelve years down-sized and my position was eliminated. After unproductive job interviews, I decided to open my own business. Over the past year, I incurred many unforeseen financial problems. I owed my family, friends and creditors over $35,000. The economy was taking a toll on my business and I just couldn't seem to make ends meet. I had to refinance and borrow against my home to support my family and struggling business. AT THAT MOMENT something significant happened in my life. I am writing to share the experience in hopes that this could change your life FOREVER. FINANCIALLY$$$!!! In mid December, I received this program in my e-mail. Six months prior to receiving this program I had been sending away for information on various business opportunities. All of the programs I received, in my opinion, were not cost effective. They were either too difficult for me to comprehend or the initial investment was too much for me to risk to see if they would work. But as I was saying, in December of 1997 I received this program. I didn't send for it, or ask for it, they just got my name off a mailing list. THANK GOODNESS FOR THAT!!! After reading it several times, to make sure I was reading it correctly. I couldn't believe my eyes! Here was a MONEY MAKING MACHINE I could start immediately without any debt. Like most of you I was still a little skeptical and a little worried about the legal aspects of it all. So I checked it out with the U.S. Post Office (1-800-725-2161 24-hrs) and they confirmed that it is indeed legal! After determining the program was LEGAL I decided "WHY NOT!?!??" Initially I sent out 10,000 e-mails. It cost me about $15 for my time on-line. The great thing about e-mail is that I don't need any for printing to send out the program, and because I also send the product (reports) by e-mail, my only expense is my time. In less than one week, I was starting to receive orders for REPORT #1. By January 13, I had received 26 orders for REPORT #1. Your goal is to "RECEIVE at least 20 ORDERS FOR REPORT #1 WITHIN 2 WEEKS. IF YOU DON'T, SEND OUT MORE PROGRAMS UNTIL YOU DO. My first step in making $50,000 in 90 days was done. By January 30, I had received 196 orders for REPORT #2. Your goal is to "RECEIVE AT LEAST 100+ ORDERS FOR REPORT #2 WITHIN 2 WEEKS. IF NOT, SEND OUT MORE PROGRAMS UNTIL YOU DO. ONCE YOU HAVE 100 ORDERS, THE REST IS EASY, RELAX, YOU WILL MAKE YOUR $50,000 GOAL." Well, I had 196 orders for REPORT #2. 96 more than I needed. So I sat back and relaxed. By March 1, of my e-mailing of 10,000, received $58,000 with more coming in every day. I paid off ALL my debts and bought a much needed new car! Please take your time to read this plan, IT WILL CHANGE YOUR LIFE FOREVER$!!! Remember, it won't work if you don't try it. This program does work, BUT you must follow it EXACTLY! Especially the rules of not trying to place your name in a different place. It won't work and you'll lose out on a lot of money! In order for this program to work, you must meet your goal of 20+ orders for REPORT #1, and 100+ orders for REPORT #2 and you will make $50,000 or more in 90 days. I AM LIVING PROOF THAT IT WORKS!!! If you choose not to participate in this program, I am sorry. It really is a great opportunity with little cost or risk to you. If you choose to participate, follow the program and you will be on your way to financial security. If you are a fellow business owner and are in financial trouble like I was, or you want to start your own business, consider this a sign. I DID! $$ Sincerely, Johnathon Rourke ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A PERSONAL NOTE FROM THE ORIGINATOR OF THIS PROGRAM: By the time you have read the enclosed program and reports, you should have concluded that such a program, and one that is legal, could not have been created by an amateur. Let me tell you a little about myself. I had a profitable business for 10 years. Then in 1979 my business began falling off. I was doing the same things that were previously successful for me, but it wasn't working. Finally, I figured it out. It wasn't me, it was the economy. Inflation and recession had replaced the stable economy that had been with us since 1945. I don't have to tell you what happened to the unemployment rate... because many of you know from first hand experience. There were more failures and bankruptcies than ever before. The middle class was vanishing. Those who knew what they were doing invested wisely and moved up. Those who did not, including those who never had anything to save or invest, were moving down into the ranks of the poor. As the saying goes, "THE RICH GET RICHER AND THE POOR GET POORER." The traditional methods of making money will never allow you to"move up" or "get rich", inflation will see to that. You have just received information that can give you financial freedom for the rest of your life, with "NO RISK" and "JUST A LITTLE BIT OF EFFORT." You can make more money in the next few months than you have ever imagined. I should also point out that I will not see a penny of this money, nor anyone else who has provided a testimonial for this program. I have retired from the program after sending thousands and thousands of programs. Follow the program EXACTLY AS INSTRUCTED. Do not change it in any way. It works exceedingly well as it is now. Remember to e-mail a copy of this exciting report to everyone you can think of. One of the people you send this to may send out 50,000...and your name will be on everyone of them! REMEMBER though, ----- the MORE YOU SEND OUT, the more potential customers you will reach. So my friend, I have given you the ideas, information, materials and opportunity to become financially independent. IT IS UP TO YOU!! NOW DO IT!! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ BEFORE YOU delete this program from your in box, as I almost did, take a little time to read it and REALLY THINK ABOUT IT. Get a pencil and figure out what could happen when YOU participate. Figure out the worst possible response and no matter how you calculate it, you will still make a lot of money! You will definitely get back what you invested. Any doubts you have will vanish when your first orders come in. $$$ IT WORKS!!! $$$ Jody Jacobs Richmond, VA ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ HERE'S HOW THIS AMAZING PROGRAM WILL MAKE YOU THOUSANDS OF DOLLAR$$$$!!!! This method of raising capital REALLY WORKS 100% EVERY TIME. I am sure that you could use up to $50,000 or more in the next 90 days. before you say "BULL... ", please read this program carefully. This is not a chain letter, but a perfectly legal money making business. As with all multi-level businesses, we build our business by recruiting new partners and selling our products. Every state in the USA allows you to recruit new multi-level business partners, and we sell and deliver a product for EVERY dollar received. YOUR ORDERS COME BY MAIL AND ARE FILLED BY E-MAIL, so you are not involved in personal selling. You do it privately in your own home, store or office. This is the EASIEST marketing plan anywhere! It is simply order filling by email! ******************************************************************* The product is informational and instructional material, keys to the secrets for everyone on how to open the doors to the magic world of E-COMMERCE , the information highway, the wave of the future ! PLAN SUMMARY: (1) You order the 4 reports listed below ($5 each) They come to you by email. (2) Save a copy of this entire letter and put your name after Report #1 and move the other names down. (3) Via the internet, access Yahoo.com or any of the other major search engines to locate hundreds of bulk email service companies (search for "bulk email") and have them send 25,000 - 50,000 emails for you about $49+) (4) Orders will come to you by postal mail - simply email them the Report they ordered. Let me ask you - isn't this about as easy as it gets? ******************************************************************* By the way there are over 50 MILLION email addresses with millions more joining the internet each year so don't worry about "running out" or "saturation". People are used to seeing and hearing the same advertisements every day on radio/TV. How many times have you received the same pizza flyers on your door? Then one day you are hungry for pizza and you order one. Same thing with this letter. I received this letter many times - then one day I decided it was time to try it. ******************************************************************* YOU CAN START TODAY - JUST DO THESE EASY STEPS: STEP #1. ORDER THE FOUR REPORTS Order the four reports shown on the list below (you can't sell them if you don't order them). -- For each report, send $5.00 CASH, the NAME & NUMBER OF THE REPORT YOU ARE ORDERING, YOUR E-MAIL ADDRESS, and YOUR NAME & RETURN ADDRESS (in case of a problem) to the person whose name appears on the list next to the report. MAKE SURE YOUR RETURN ADDRESS IS ON YOUR ENVELOPE IN CASE OF ANY MAIL PROBLEMS! Within a few days you will receive, by e-mail, each of the four reports. Save them on your computer so you can send them to the 1,000's of people who will order them from you. STEP #2. ADD YOUR MAILING ADDRESS TO THIS LETTER a. Look below for the listing of the four reports. b. After you've ordered the four reports, delete the name and address under REPORT #4. This person has made it through the cycle. c. Move the name and address under REPORT #3 down to REPORT #4. d. Move the name and address under REPORT #2 down to REPORT #3. e. Move the name and address under REPORT #1 down to REPORT #2. f. Insert your name/address in the REPORT #1 position. Please make sure you COPY ALL INFORMATION, every name and address, ACCURATELY! STEP #3. Take this entire letter, including the modified list of names, and save it to your computer. Make NO changes to these instructions. Now you are ready to use this entire email to send by email to prospects. Report #1 will tell you how to download bulk email software and email addresses so you can send it out to thousands of people while you sleep! Remember that 50,000+ new people are joining the internet every month. Your cost to participate in this is practically nothing (surely you can afford $20 and initial bulk mailing cost). You obviously already have a computer and an Internet connection and e-mail is FREE! There are two primary methods of building your downline: METHOD #1: SENDING BULK E-MAIL Let's say that you decide to start small, just to see how it goes, and we'll assume you and all those involved email out only 2,000 programs each. Let's also assume that the mailing receives a 0.5% response. The response could be much better. Also, many people will email out hundreds of thousands of programs instead of 2,000 (Why stop at 2000?). But continuing with this example, you send out only 2,000 programs. With a 0.5% response, that is only 10 orders for REPORT #1. Those 10 people respond by sending out 2,000 programs each for a total of 20,000. Out of those 0.5%, 100 people respond and order REPORT #2. Those 100 mail out 2,000 programs each for a total of 200,000. The 0.5% response to that is 1,000 orders for REPORT #3. Those 1,000 send out 2,000 programs each for a 2,000,000 total. The 0.5% response to that is 10,000 orders for REPORT #4. That's 10,000 $5 bills for you. CASH!!! Your total income in this example is $50 + $500 + $5,000 + $50,000 for a total of $55,550!!! REMEMBER FRIEND, THIS IS ASSUMING 1,990 OUT OF THE 2,000 PEOPLE YOU MAIL TO WILL DO ABSOLUTELY NOTHING AND TRASH THIS PROGRAM! DARE TO THINK FOR A MOMENT WHAT WOULD HAPPEN IF EVERYONE, OR HALF SENT OUT 100,000 PROGRAMS INSTEAD OF 2,000. Believe me, many people will do just that, and more! METHOD #2 - PLACING FREE ADS ON THE INTERNET Advertising on the internet is very, very inexpensive, and there are HUNDREDS of FREE places to advertise. Let's say you decide to start small just to see how well it works. Assume your goal is to get ONLY 10 people to participate on your first level. (Placing a lot of FREE ads on the Internet will EASILY get a larger response.) Also assume that everyone else in YOUR ORGANIZATION gets ONLY 10 downline members. Look how this small number accumulates to achieve the STAGGERING results below: 1st level--your first 10 send you $5 ..............................$50 2nd level--10 members from those 10 ($5 x 100)...........$500 3rd level--10 members from those 100 ($5 x 1,000).......$5,000 4th level--10 members from those 1,000 ($5 x 10,000)..$50,000 $$$$$$ THIS TOTALS -----------------------------------$55,550 $$$$$$ AMAZING ISN'T IT? Remember friends, this assumes that the people who participate only recruit 10 people each. Think for a moment what would happen if they got 20 people to participate! Most people get 100's of participants and many will continue to work this program, sending out programs WITH YOUR NAME ON THEM for years! THINK ABOUT IT! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ People are going to get emails about this plan from you or somebody else and many will work this plan - the question is - Don't you want your name to be on the emails they will send out? * * * DON'T MISS OUT!!! * * * JUST TRY IT ONCE!!! * * * * * * SEE WHAT HAPPENS!!! * * * YOU'LL BE AMAZED!!! * * * ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ALWAYS PROVIDE SAME-DAY SERVICE ON ALL ORDERS! This will guarantee that the e-mail THEY send out with YOUR name and address on it will be prompt because they can't advertise until they receive the report! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ GET STARTED TODAY: PLACE YOUR ORDER FOR THE FOUR REPORTS NOW. Notes: -- ALWAYS SEND $5 CASH (U.S. CURRENCY) FOR EACH REPORT. CHECKS NOT ACCEPTED. Make sure the cash is concealed by wrapping it in two sheets of paper. On one of those sheets of paper write: (a) the number & name of the report you are ordering (b) your e-mail address, and (c) your name & postal address. ---------------------------------------------------------------------- REPORT #1 "The Insider's Guide to Advertising for Free on the Internet" ORDER REPORT #1 FROM: MAIN STREET P O BOX 598 BOLIVAR TN 38008-0598 NOTE: I and every member below are dedicated at helping you with this program so it will work for you also. TRY US! ------------------------------------------------------------------------ REPORT #2 "The Insider's Guide to Sending Bulk E-mail on the Internet" ORDER REPORT #2 FROM: MARCUS MILLER 918 KNOLLFIELD WAY SAN JOSE CA 9959136 ------------------------------------------------------------------------ REPORT #3 "The Secrets to Multilevel Marketing on the Internet" ORDER REPORT #3 FROM: LANCE DENTON a.k.a (Debtfighter) 100 WILDWOOD RD ROCKPORT TX 78382 ------------------------------------------------------------------------- REPORT #4 "How to become a Millionaire utilizing the Power of Multilevel Marketing and the Internet" ORDER REPORT #4 FROM: MELISSA HOGENMILLER 3709 MONHEIM RD CONOVER WI 54519 -------------------------------------------------------------------------- ******* TIPS FOR SUCCESS ******* TREAT THIS AS YOUR BUSINESS! Be prompt, professional, and follow the directions accurately. -- Send for the four reports IMMEDIATELY so you will have them when the orders start coming in because: When you receive a $5 order, you MUST send out the requested product/report. It is required for this to be a legal business and they need the reports to send out their letters (with your name on them!) -- ALWAYS PROVIDE SAME-DAY SERVICE ON THE ORDERS YOU RECEIVE. -- Be patient and persistent with this program - If you follow the instructions exactly - results WILL FOLLOW. $$$$ ******* YOUR SUCCESS GUIDELINES ******* Follow these guidelines to guarantee your success: If you don't receive 20 orders for REPORT #1 within two weeks, continue advertising or sending e-mails until you do. Then, a couple of weeks later you should receive at least 100 orders for REPORT#2. If you don't, continue advertising or sending e-mails until you do. Once you have received 100 or more orders for REPORT #2, YOU CAN RELAX, because the system is already working for you, and the cash will continue to roll in! THIS IS IMPORTANT TO REMEMBER: Every time your name is moved down on the list, you are placed in front of a DIFFERENT report. You can KEEP TRACK of your PROGRESS by watching which report people are ordering from you. To generate more income, simply send another batch of e-mails or continue placing ads and start the whole process again! There is no limit to the income you will generate from this business! Before you make your decision as to whether or not you participate in this program. Please answer one question. ARE YOU HAPPY WITH YOUR PRESENT INCOME OR JOB? If the answer is no, then please look at the following facts about this super simple MLM program: 1. NO face to face selling, NO meetings, NO inventory! NO Telephone calls, NO big cost to start!, NOthing to learn, NO skills needed! (Surely you know how to send email?) 2. No equipment to buy - you already have a computer and internet connection - so you have everything you need to fill orders! 3. You are selling a product which does NOT COST ANYTHING TO PRODUCE OR SHIP! (Emailing copies of the reports is FREE!) 4. All of your customers pay you in CA$H! This program will change your LIFE FOREVER!! Look at the potential for you to be able to quit your job and live a life of luxury you could only dream about! Imagine getting out of debt and buying the car and home of your dreams and being able to work a super-high paying leisurely easy business from home! $$$ FINALLY MAKE SOME DREAMS COME TRUE! $$$ ACT NOW! Take your first step toward achieving financial independence. Order the reports and follow the program outlined above-- SUCCESS will be your reward. Thank you for your time and consideration. PLEASE NOTE: If you need help with starting a business, registering a business name, learning how income tax is handled, etc., contact your local office of the Small Business Administration (a Federal Agency) 1-800-827-5722 for free help and answers to questions. Also, the Internal Revenue Service offers free help via telephone and free seminars about business tax requirements. Your earnings are highly dependent on your activities and advertising. The information contained on this site and in the report constitutes no guarantees stated nor implied. In the event that it is determined that this site or report constitutes a guarantee of any kind, that guarantee is now void. The earnings amounts listed on this site and in the report are estimates only. If you have any questions of the legality of this program, contact the Office of Associate Director for Marketing Practices, Federal Trade Commission, Bureau of Consumer Protection in Washington, DC. ================================================ Under Bill s.1618 TITLE III passed by the 105th US Congress this letter cannot be considered spam as long as the sender includes contact information and a method of removal. This is a one time e-mail transmission. No request for removal is necessary. --- From owner-fluorpro@hgmp.mrc.ac.uk Sat May 27 22:26:08 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id D46B117B43; Sat, 27 May 2000 22:26:06 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id C1A5717B0B; Sat, 27 May 2000 22:26:05 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 11C7D415D5 for ; Sat, 27 May 2000 22:26:04 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA20133 for fluorpro-list@hgmp.mrc.ac.uk; Sat, 27 May 2000 22:26:04 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "brulinodia@katamail.com" X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: GFP in Yeast Date: Sat, 27 May 2000 23:26:06 +0200 Lines: 14 Message-ID: <39303D6E.1F2ACC6F@katamail.com> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: serv1.albacom.net 959462760 23611 213.255.30.158 (27 May 2000 21:26:00 GMT) X-Complaints-To: abuse@albacom.net X-Mailer: Mozilla 4.51 [it] (Win98; I) X-Accept-Language: it To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Hi there does anybody knows ig GFF works in Yeast? Better, does anybody knows if fluorescence leaks out from the cell wall? Thanks in advance Guido Cimoli Dpt. Experimental Oncology Natl.Cancer.Inst. Genoa Italy email: luther@hp380.ist.unige.it From owner-fluorpro@hgmp.mrc.ac.uk Sun May 28 21:37:17 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C88F717A9F; Sun, 28 May 2000 21:37:15 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id B320A17A4E; Sun, 28 May 2000 21:37:14 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 2FAC0415CA for ; Sun, 28 May 2000 21:37:14 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id VAA15648 for fluorpro-list@hgmp.mrc.ac.uk; Sun, 28 May 2000 21:37:13 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: mark.4902@belgium.com X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Work From Home! 2-5k Per Week! Date: 28 May 2000 21:37:12 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 59 Message-ID: <200005281924.VAA30920@relay.nre.at> X-Trace: niobium.hgmp.mrc.ac.uk 959546233 15646 193.62.192.80 (28 May 2000 20:37:13 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id DA33817A4E; Sun, 28 May 2000 21:37:10 +0100 (BST) X-Received: from relay.nre.at (relay.nre.at [195.58.184.59]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 33746415CA for ; Sun, 28 May 2000 21:37:09 +0100 (BST) X-Received: from unknown (04-177.010.popsite.net [208.178.88.177]) by relay.nre.at (8.9.3/8.8.7) with SMTP id VAA30920; Sun, 28 May 2000 21:24:19 +0200 X-To: fd09la@jkflap.com To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk LOOKING FOR GEMS! It's So Simple To Earn $2,000 - $5,000 Per Week Nowadays... We're searching for only 10 elite individuals with the work ethic necessary to generate a cash-flow for themselves of $2,000 - $5,000 per week, and to increase that to over $20,000 per month, in as little as four to six months. And you know what? If you really have a burning desire and commitment, we guarantee you that you'll reach this explosive income! Can you read a short script to our qualified leads, and then turn the interested prospects over to our electronic sales medium? (you will not be required to do any selling.) Do you have the self-discipline to ignore the TV for a couple of hours per day? Are you looking for a legitimate home-based business opportunity, that is not multi-level marketing, or a chain-letter scheme? If you would like to build an amazing income that will grow lightning-fast and have you profit $1,000.00 every time only one prospect makes a purchase, then this is for you! You can build the business under our guidance and support without having to attend meetings or sell people things they don't need. Call NOW our TOLL FREE, PRE-RECORDED Message:1-800-587-9046 Ext. 8379 We market a real product, that pays real commissions to you,$1,000.00 per sale, just for making the initial contacts. With our turn-key lead generation systems you'll always talk to people who actually WANT to talk to you. You have nothing to lose, there's no risk involved, nor is there any obligation whatsoever, and you may be qualified to earn thousands of extra dollars per month! So call now! The call is FREE, and there is absolutely no obligation, So what have you got to lose? Call Toll Free 1-800-587-9064 Ext. 8379 P.S. You literally have a once-in-a-lifetime opportunity to GET INVOLVED NOW! Don't let this one go by. You have absolutely nothing to lose! This could be the most fascinating and profitable business of your life! Please, serious inquiries only. We apologize for any inconvenience hitting the delete key has caused if you did not wish to receive this message. This message is being sent to you in compliance with the proposed Federal legislation for commercial e-mail (S.1618 - SECTION 301). "Pursuant to Section 301, Paragraph (a)(2)(C) of S. 1618, To be removed from our mailing list please send an email to: remove.me.pls@HotPOP.com and place remove in the subject Thank you --- From owner-fluorpro@hgmp.mrc.ac.uk Tue May 30 18:37:04 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 9BBA317B2D; Tue, 30 May 2000 18:37:02 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 49D4317B24; Tue, 30 May 2000 18:37:01 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id BA1D9415D5 for ; Tue, 30 May 2000 18:37:00 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id SAA26304 for fluorpro-list@hgmp.mrc.ac.uk; Tue, 30 May 2000 18:36:59 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: patricia6123_9@mpg-net.de X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: How have you been? Do you remember me? Date: 30 May 2000 18:36:59 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 529 Message-ID: <200005310140.BAA24533@ns1.rtm.net.my> Mime-Version: 1.0 Content-Type: text/plain; charset=unknown-8bit X-Trace: niobium.hgmp.mrc.ac.uk 959708219 26302 193.62.192.80 (30 May 2000 17:36:59 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 75C3517B24; Tue, 30 May 2000 18:36:57 +0100 (BST) X-Received: from ns1.rtm.net.my (ns1.rtm.net.my [202.190.233.3]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id D2E29415D5 for ; Tue, 30 May 2000 18:36:47 +0100 (BST) X-Received: from monorailpc (ppp-54.tnt-1.ocl.smartworld.net [64.38.40.54]) by ns1.rtm.net.my (8.8.8/8.7.1) with SMTP id BAA24533; Wed, 31 May 2000 01:40:32 GMT X-To: patricia6123_9@mpg-net.de To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk So, how have you been? Perhaps you remember holding previous conversations regarding business and money making opportunities. I did not send this to you in error! You said: "If only I could find an easier way to make a higher income" and "If I had more money, I could spend more time with my Family, and less time at work" and "I sure could use more money so I could pay off my bills once and for all!" and "I would love to get involved in a business in which will generate money while I am not at work (like a Gas Pump)." Dear Friend, There is a possibility that we haven't met, but you were chosen by someone to recieve this E-Mail. Please, please, print this off and readthoroughly. Be sure that you don't miss any of the points outlined. Then put it down, and then read it again. I am sending you a whole lot of information in which you might not understand the first time you read it. If you don't believe this program will work for you, send it to 10-20 of your closest friends (in which you trust deeply), and ask them what they think? This really works! Have the faith, don't miss this opportunity, get involved also, and it will work for you as it does for us!!!!! Due to the popularity of this letter on the Internet, A MAJOR NIGHTLY NEWS Program recently dedicated entire show to the investigation of the program described below to see if it really can make people money. The show also investigated whether or not the program was legal. Their findings proved that there are absolutely no laws prohibiting the participation in the program. This has helped to show people that this is a simple, harmless and fun way to make some extra money at home. The results have been truly remarkable. So many people are participating that those involved are doing much better than ever before. Since everyone makes more as more people try it out, its been very exciting. You will understand only if you get involved! ********* THE ENTIRE PLAN IS HERE BELOW ********* *** Print This Now For Future Reference *** $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ If you would like to make AT LEAST $50,000 in less than 90 days! If not, forward this to someone who would like to make this kind of money....... It works (like designed) but only for those who follow it to the letter! Please read this program...THEN READ IT AGAIN!! $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ THIS IS A LEGITIMATE, LEGAL, MONEY MAKING OPPORTUNITY!! It does NOT require you to come into contact with people or make or take any telephone calls. Just follow the instructions, and you will make money. This simplified e-mail marketing program works perfectly 100% EVERY TIME! E-mail is the sales tool of the future. Take advantage of this virtually free method of advertising NOW!!! The longer you wait, the more people will be doing business using email. Get your piece of this action!!! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Hello - My name is Johnathon Rourke, I'm from Rhode Island. The enclosed information is something I almost let slip through my fingers. Fortunately, sometime later I re-read everything and gave some thought and study to it. Two years ago, the corporation I worked for the past twelve years down-sized and my position was eliminated. After unproductive job interviews, I decided to open my own business. Over the past year, I incurred many unforeseen financial problems. I owed my family, friends and creditors over $35,000. The economy was taking a toll on my business and I just couldn't seem to make ends meet. I had to refinance and borrow against my home to support my family and struggling business. AT THAT MOMENT something significant happened in my life. I am writing to share the experience in hopes that this could change your life FOREVER. FINANCIALLY$$$!!! In mid December, I received this program in my e-mail. Six months prior to receiving this program I had been sending away for information on various business opportunities. All of the programs I received, in my opinion, were not cost effective. They were either too difficult for me to comprehend or the initial investment was too much for me to risk to see if they would work. But as I was saying, in December of 1997 I received this program. I didn't send for it, or ask for it, they just got my name off a mailing list. THANK GOODNESS FOR THAT!!! After reading it several times, to make sure I was reading it correctly. I couldn't believe my eyes! Here was a MONEY MAKING MACHINE I could start immediately without any debt. Like most of you I was still a little skeptical and a little worried about the legal aspects of it all. So I checked it out with the U.S. Post Office (1-800-725-2161 24-hrs) and they confirmed that it is indeed legal! After determining the program was LEGAL I decided "WHY NOT!?!??" Initially I sent out 10,000 e-mails. It cost me about $15 for my time on-line. The great thing about e-mail is that I don't need any for printing to send out the program, and because I also send the product (reports) by e-mail, my only expense is my time. In less than one week, I was starting to receive orders for REPORT #1. By January 13, I had received 26 orders for REPORT #1. Your goal is to "RECEIVE at least 20 ORDERS FOR REPORT #1 WITHIN 2 WEEKS. IF YOU DON'T, SEND OUT MORE PROGRAMS UNTIL YOU DO. My first step in making $50,000 in 90 days was done. By January 30, I had received 196 orders for REPORT #2. Your goal is to "RECEIVE AT LEAST 100+ ORDERS FOR REPORT #2 WITHIN 2 WEEKS. IF NOT, SEND OUT MORE PROGRAMS UNTIL YOU DO. ONCE YOU HAVE 100 ORDERS, THE REST IS EASY, RELAX, YOU WILL MAKE YOUR $50,000 GOAL." Well, I had 196 orders for REPORT #2. 96 more than I needed. So I sat back and relaxed. By March 1, of my e-mailing of 10,000, received $58,000 with more coming in every day. I paid off ALL my debts and bought a much needed new car! Please take your time to read this plan, IT WILL CHANGE YOUR LIFE FOREVER$!!! Remember, it won't work if you don't try it. This program does work, BUT you must follow it EXACTLY! Especially the rules of not trying to place your name in a different place. It won't work and you'll lose out on a lot of money! In order for this program to work, you must meet your goal of 20+ orders for REPORT #1, and 100+ orders for REPORT #2 and you will make $50,000 or more in 90 days. I AM LIVING PROOF THAT IT WORKS!!! If you choose not to participate in this program, I am sorry. It really is a great opportunity with little cost or risk to you. If you choose to participate, follow the program and you will be on your way to financial security. If you are a fellow business owner and are in financial trouble like I was, or you want to start your own business, consider this a sign. I DID! $$ Sincerely, Johnathon Rourke ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A PERSONAL NOTE FROM THE ORIGINATOR OF THIS PROGRAM: By the time you have read the enclosed program and reports, you should have concluded that such a program, and one that is legal, could not have been created by an amateur. Let me tell you a little about myself. I had a profitable business for 10 years. Then in 1979 my business began falling off. I was doing the same things that were previously successful for me, but it wasn't working. Finally, I figured it out. It wasn't me, it was the economy. Inflation and recession had replaced the stable economy that had been with us since 1945. I don't have to tell you what happened to the unemployment rate... because many of you know from first hand experience. There were more failures and bankruptcies than ever before. The middle class was vanishing. Those who knew what they were doing invested wisely and moved up. Those who did not, including those who never had anything to save or invest, were moving down into the ranks of the poor. As the saying goes, "THE RICH GET RICHER AND THE POOR GET POORER." The traditional methods of making money will never allow you to"move up" or "get rich", inflation will see to that. You have just received information that can give you financial freedom for the rest of your life, with "NO RISK" and "JUST A LITTLE BIT OF EFFORT." You can make more money in the next few months than you have ever imagined. I should also point out that I will not see a penny of this money, nor anyone else who has provided a testimonial for this program. I have retired from the program after sending thousands and thousands of programs. Follow the program EXACTLY AS INSTRUCTED. Do not change it in any way. It works exceedingly well as it is now. Remember to e-mail a copy of this exciting report to everyone you can think of. One of the people you send this to may send out 50,000...and your name will be on everyone of them! REMEMBER though, ----- the MORE YOU SEND OUT, the more potential customers you will reach. So my friend, I have given you the ideas, information, materials and opportunity to become financially independent. IT IS UP TO YOU!! NOW DO IT!! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ BEFORE YOU delete this program from your in box, as I almost did, take a little time to read it and REALLY THINK ABOUT IT. Get a pencil and figure out what could happen when YOU participate. Figure out the worst possible response and no matter how you calculate it, you will still make a lot of money! You will definitely get back what you invested. Any doubts you have will vanish when your first orders come in. $$$ IT WORKS!!! $$$ Jody Jacobs Richmond, VA ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ HERE'S HOW THIS AMAZING PROGRAM WILL MAKE YOU THOUSANDS OF DOLLAR$$$$!!!! This method of raising capital REALLY WORKS 100% EVERY TIME. I am sure that you could use up to $50,000 or more in the next 90 days. before you say "BULL... ", please read this program carefully. This is not a chain letter, but a perfectly legal money making business. As with all multi-level businesses, we build our business by recruiting new partners and selling our products. Every state in the USA allows you to recruit new multi-level business partners, and we sell and deliver a product for EVERY dollar received. YOUR ORDERS COME BY MAIL AND ARE FILLED BY E-MAIL, so you are not involved in personal selling. You do it privately in your own home, store or office. This is the EASIEST marketing plan anywhere! It is simply order filling by email! ******************************************************************* The product is informational and instructional material, keys to the secrets for everyone on how to open the doors to the magic world of E-COMMERCE , the information highway, the wave of the future ! PLAN SUMMARY: (1) You order the 4 reports listed below ($5 each) They come to you by email. (2) Save a copy of this entire letter and put your name after Report #1 and move the other names down. (3) Via the internet, access Yahoo.com or any of the other major search engines to locate hundreds of bulk email service companies (search for "bulk email") and have them send 25,000 - 50,000 emails for you about $49+) (4) Orders will come to you by postal mail - simply email them the Report they ordered. Let me ask you - isn't this about as easy as it gets? ******************************************************************* By the way there are over 50 MILLION email addresses with millions more joining the internet each year so don't worry about "running out" or "saturation". People are used to seeing and hearing the same advertisements every day on radio/TV. How many times have you received the same pizza flyers on your door? Then one day you are hungry for pizza and you order one. Same thing with this letter. I received this letter many times - then one day I decided it was time to try it. ******************************************************************* YOU CAN START TODAY - JUST DO THESE EASY STEPS: STEP #1. ORDER THE FOUR REPORTS Order the four reports shown on the list below (you can't sell them if you don't order them). -- For each report, send $5.00 CASH, the NAME & NUMBER OF THE REPORT YOU ARE ORDERING, YOUR E-MAIL ADDRESS, and YOUR NAME & RETURN ADDRESS (in case of a problem) to the person whose name appears on the list next to the report. MAKE SURE YOUR RETURN ADDRESS IS ON YOUR ENVELOPE IN CASE OF ANY MAIL PROBLEMS! Within a few days you will receive, by e-mail, each of the four reports. Save them on your computer so you can send them to the 1,000's of people who will order them from you. STEP #2. ADD YOUR MAILING ADDRESS TO THIS LETTER a. Look below for the listing of the four reports. b. After you've ordered the four reports, delete the name and address under REPORT #4. This person has made it through the cycle. c. Move the name and address under REPORT #3 down to REPORT #4. d. Move the name and address under REPORT #2 down to REPORT #3. e. Move the name and address under REPORT #1 down to REPORT #2. f. Insert your name/address in the REPORT #1 position. Please make sure you COPY ALL INFORMATION, every name and address, ACCURATELY! STEP #3. Take this entire letter, including the modified list of names, and save it to your computer. Make NO changes to these instructions. Now you are ready to use this entire email to send by email to prospects. Report #1 will tell you how to download bulk email software and email addresses so you can send it out to thousands of people while you sleep! Remember that 50,000+ new people are joining the internet every month. Your cost to participate in this is practically nothing (surely you can afford $20 and initial bulk mailing cost). You obviously already have a computer and an Internet connection and e-mail is FREE! There are two primary methods of building your downline: METHOD #1: SENDING BULK E-MAIL Let's say that you decide to start small, just to see how it goes, and we'll assume you and all those involved email out only 2,000 programs each. Let's also assume that the mailing receives a 0.5% response. The response could be much better. Also, many people will email outhundreds of thousands of programs instead of 2,000 (Why stop at 2000?). But continuing with this example, you send out only 2,000 programs. With a 0.5% response, that is only 10 orders for REPORT #1. Those 10 people respond by sending out 2,000 programs each for a total of 20,000. Out of those 0.5%, 100 people respond and order REPORT #2. Those 100 mail out 2,000 programs each for a total of 200,000. The 0.5% response to that is 1,000 orders for REPORT #3. Those 1,000 send out 2,000 programs each for a 2,000,000 total. The 0.5% response to that is 10,000 orders for REPORT #4. That's 10,000 $5 bills for you. CASH!!! Your total income in this example is $50 + $500 + $5,000 + $50,000 for a total of $55,550!!! REMEMBER FRIEND, THIS IS ASSUMING 1,990 OUT OF THE 2,000 PEOPLE YOU MAIL TO WILL DO ABSOLUTELY NOTHING AND TRASH THIS PROGRAM! DARE TO THINK FOR A MOMENT WHAT WOULD HAPPEN IF EVERYONE, OR HALF SENT OUT 100,000 PROGRAMS INSTEAD OF 2,000. Believe me, many people will do just that, and more! METHOD #2 - PLACING FREE ADS ON THE INTERNET Advertising on the internet is very, very inexpensive, and there are HUNDREDS of FREE places to advertise. Let's say you decide to start small just to see how well it works. Assume your goal is to get ONLY 10 people to participate on your first level. (Placing a lot of FREE ads on the Internet will EASILY get a larger response.) Also assume that everyone else in YOUR ORGANIZATION gets ONLY 10 downline members. Look how this small number accumulates to achieve the STAGGERING results below: 1st level--your first 10 send you $5 .....................$50 2nd level--10 members from those 10 ($5 x 100)...........$500 3rd level--10 members from those 100 ($5 x 1,000)......$5,000 4th level--10 members from those 1,000 ($5 x 10,000)..$50,000 $$$$$$ THIS TOTALS -----------------------------------$55,550 $$$$$$ AMAZING ISN'T IT? Remember friends, this assumes that the people who participate only recruit 10 people each. Think for a moment what would happen if they got 20 people to participate! Most people get 100's of participants and many will continue to work this program, sending out programs WITH YOUR NAME ON THEM for years! THINK ABOUT IT! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ People are going to get emails about this plan from you or somebody else and many will work this plan - the question is - Don't you want your name to be on the emails they will send out? * * * DON'T MISS OUT!!! * * * JUST TRY IT ONCE!!! * * * * * SEE WHAT HAPPENS!!! *** YOU'LL BE AMAZED!!!* * ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ALWAYS PROVIDE SAME-DAY SERVICE ON ALL ORDERS! This will guarantee that the e-mail THEY send out with YOUR name and address on it will be prompt because they can't advertise until they receive the report! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ GET STARTED TODAY: PLACE YOUR ORDER FOR THE FOUR REPORTS NOW. Notes: -- ALWAYS SEND $5 CASH (U.S. CURRENCY) FOR EACH REPORT. CHECKS NOT ACCEPTED. Make sure the cash is concealed by wrapping it in two sheets of paper. On one of those sheets of paper write: (a) the number & name of the report you are ordering (b) your e-mail address, and (c) your name & postal address. REPORT #1b "The Insider's Guide to Advertising for Free on the Internet" ORDER REPORT #1 FROM: Chaplain Jeff Congdon P.O. Box 456 Midway, TX 75852 NOTE: I and every member below are dedicated at help you with this program so it will work for you also. TRY US! REPORT #2b "The Insider's Guide to Sending Bulk E-mail on the Internet" ORDER REPORT #2 FROM: Bruce Rutherford 174 McKeen Pl. Penticton, B.C. V2A 6W1 REPORT #3b "The Secrets to Multilevel Marketing on the Internet" ORDER REPORT #3 FROM: Lance Denton 100 Wildwood Rd. Rockport, TX 78382 REPORT #4b "How to become a Millionaire utilizing the Power of Multilevel Marketing and the Internet" ORDER REPORT #4 FROM: Melissa Hogenmiller 3709 Monheim Rd. Convoer, Wis. 54519 ******* TIPS FOR SUCCESS ******* TREAT THIS AS YOUR BUSINESS! Be prompt, professional, and follow the directions accurately. -- Send for the four reports IMMEDIATELY so you will have them when the orders start coming in because: When you receive a $5 order, you MUST send out the requested product/report. It is required for this to be a legal business and they need the reports to send out their letters (with your name on them!) -- ALWAYS PROVIDE SAME-DAY SERVICE ON THE ORDERS YOU RECEIVE. -- Be patient and persistent with this program - If you follow the instructions exactly - results WILL FOLLOW. $$$$ ******* YOUR SUCCESS GUIDELINES ******* Follow these guidelines to guarantee your success: If you don't receive 20 orders for REPORT #1 within two weeks, continue advertising orsending e-mails until you do. Then, a couple of weeks later you should receive at least 100 orders for REPORT#2. If you don't, continue advertising or sending e-mails until you do. Once you have received 100 or more orders for REPORT #2, YOU CAN RELAX, because the system is already working for you, and the cash will continue to roll in! THIS IS IMPORTANT TO REMEMBER: Every time your name is moved down on the list, you are placed in front of a DIFFERENT report. You can KEEP TRACK of your PROGRESS by watching which report people are ordering from you. To generate more income, simply send another batch of e-mails or continue placing ads and start the whole process again! There is no limit to the income you will generate from this business! Before you make your decision as to whether or not you participate in this program. Please answer one question. ARE YOU HAPPY WITH YOUR PRESENT INCOME OR JOB? If the answer is no, then please look at the following facts about this super simple MLM program: 1. NO face to face selling, NO meetings, NO inventory! NO Telephone calls, NO big cost to start!, NOthing to learn, NO skills needed! (Surely you know how to send email?) 2. No equipment to buy - you already have a computer and internet connection - so you have everything you need to fill orders! 3. You are selling a product which does NOT COST ANYTHING TO PRODUCE OR SHIP! (Emailing copies of the reports is FREE!) 4. All of your customers pay you in CA$H! This program will change your LIFE FOREVER!! Look at the potential for you to be able to quit your job and live a life of luxury you could only dream about! Imagine getting out of debt and buying the car and home of your dreams and being able to work a super-high paying leisurely easy business from home! $$$ FINALLY MAKE SOME DREAMS COME TRUE! $$$ ACT NOW! Take your first step toward achieving financial independence. Order the reports and follow the program outlined above-- SUCCESS will be your reward. Thank you for your time and consideration. PLEASE NOTE: If you need help with starting a business, registering a business name, learning how income tax is handled, etc., contact your local office of the Small Business Administration (a Federal Agency) 1-800-827-5722 for free help and answers to questions. Also, the Internal Revenue Service offers free help via telephone and free seminars about business tax reuirements. Your earnings are highly dependent on your activities and advertising. The information contained on this site and in the report constitutes no guarantees stated nor implied. In the event that it is determined that this site or report constitutes a guarantee of any kind, that guarantee is now void. The earnings amounts listed on this site and in the report are estimates only. If you have any questions of the legality of this program, contact the Office of Associate Director for Marketing Practices, Federal Trade Commission, Bureau of Consumer Protection in Washington, DC. ================================================ Under Bill s.1618 TITLE III passed by the 105th US Congress this letter cannot be considered spam as long as the sender includes contact information and a method of removal. This is a one time e-mail transmission. No request for removal is necessary. ________________________________________________________________________ --- From owner-fluorpro@hgmp.mrc.ac.uk Wed May 31 14:01:33 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3F9CC17B31; Wed, 31 May 2000 14:01:31 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 1999817B2C; Wed, 31 May 2000 14:01:28 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 61083415DC for ; Wed, 31 May 2000 14:01:28 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA19558 for fluorpro-list@hgmp.mrc.ac.uk; Wed, 31 May 2000 14:01:27 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: RN@AMI.DK ("Rikke Nikolajsen") X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: catecholamine fluorescence and pH Date: 31 May 2000 14:01:26 +0100 Organization: =?ISO-8859-1?Q?Arbejdsmilj=F8instituttet?= Lines: 20 Message-ID: <20000531130123.C6310415DC@mercury.hgmp.mrc.ac.uk> Mime-Version: 1.0 Content-Type: text/plain; charset=ISO-8859-1 Content-Transfer-Encoding: Quoted-printable X-Trace: niobium.hgmp.mrc.ac.uk 959778087 19555 193.62.192.80 (31 May 2000 13:01:27 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id D5EA317B2C; Wed, 31 May 2000 14:01:24 +0100 (BST) X-Received: from mail.net.uni-c.dk (mail.net.uni-c.dk [130.226.1.3]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id C6310415DC for ; Wed, 31 May 2000 14:01:23 +0100 (BST) X-Received: (qmail 6678 invoked from network); 31 May 2000 13:01:22 -0000 X-Received: from ami.dk (130.226.238.100) by mail.net.uni-c.dk with SMTP; 31 May 2000 13:01:22 -0000 X-Received: from AMILIA/SpoolDir by AMI.DK (Mercury 1.47); 31 May 00 15:00:41 +0100 X-Received: from SpoolDir by AMILIA (Mercury 1.47); 31 May 00 15:00:24 +0100 X-To: fluorpro@net.bio.net X-Priority: normal X-Mailer: Pegasus Mail for Win32 (v3.01d) To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Hello - I am not quite sure, this is the right newsgroup to ask this question in - but I'm more or less desperate..... I would like to know how the fluorescence spectra of the catecholamines adrenaline/ephinephrine and noradrenaline/norepinephrine) change with pH . The compounds seem to decompose in even slightly basic media (e.g. pH=3D8.5) - the fluorescence vanishes. Is this due to oxidation of phenoles??? =96 or is it another cause? Will you under any circumstances be able to measure catecholamine fluorescence in basic solution? Furthermore, does anybody know where the catecholamine's chemical and fluorescence properties might be described in detail? Thanks in advance Rikke --- From owner-fluorpro@hgmp.mrc.ac.uk Wed May 31 22:52:29 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5378417AC4; Wed, 31 May 2000 22:52:28 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id DB84517A9F; Wed, 31 May 2000 22:52:26 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 44070415CA for ; Wed, 31 May 2000 22:52:26 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA04694 for fluorpro-list@hgmp.mrc.ac.uk; Wed, 31 May 2000 22:52:25 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: bludin@lis.ch (Beat Ludin) X-Newsgroups: bionet.molbio.proteins.fluorescent Subject: Re: catecholamine fluorescence and pH Date: 31 May 2000 22:52:24 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 41 Message-ID: Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" Content-Transfer-Encoding: quoted-printable X-Trace: niobium.hgmp.mrc.ac.uk 959809945 4692 193.62.192.80 (31 May 2000 21:52:25 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id C445817A9F; Wed, 31 May 2000 22:52:22 +0100 (BST) X-Received: from gamma.dynaweb.de (unknown [195.127.244.2]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 89322415CA for ; Wed, 31 May 2000 22:52:21 +0100 (BST) X-Received: from [195.24.82.252] ([195.24.82.252]) by gamma.dynaweb.de (8.9.1/8.9.1) with ESMTP id XAA06921 for ; Wed, 31 May 2000 23:51:56 +0200 (CEST) X-Sender: lis3@pop.lis.ch X-To: GFP Mailing Llist To: fluorpro@hgmp.mrc.ac.uk Sender: owner-fluorpro@hgmp.mrc.ac.uk Precedence: bulk Sorry, I can't answer your questions. I'm just curious, have you=20 checked whether the fluorescence returns (immediately) upon=20 neutralization? The would suggest a simple de-protonation. Cheers, Beat >Hello - I am not quite sure, this is the right newsgroup to ask this >question in - but I'm more or less desperate..... > >I would like to know how the fluorescence spectra of the catecholamines >adrenaline/ephinephrine and noradrenaline/norepinephrine) change with >pH . > >The compounds seem to decompose in even slightly basic media (e.g. >pH=3D8.5) - the fluorescence vanishes. Is this due to oxidation of >phenoles??? =96 or is it another cause? Will you under any circumstances >be able to measure catecholamine fluorescence in basic solution? > >Furthermore, does anybody know where the catecholamine's chemical >and fluorescence properties might be described in detail? > >Thanks in advance >Rikke > > >--- LIFE IMAGING SERVICES =20 - visit our web site at http://www.lis.ch +----------------------------------------------------------- | Dr. B. Ludin | Life Imaging Services fon ++41 (0)79 235 7154 | Muehletalweg 22 fax ++41 (0)860 622 963 160 | CH-4600 Olten beat.ludin@lis.ch | Switzerland http://www.lis.ch ---