From bhawana from chembiotek.com Thu Aug 13 06:01:43 2009 From: bhawana from chembiotek.com (bhawana@chembiotek.com) Date: Thu Aug 13 12:02:01 2009 Subject: [Cell-biology] need your help Message-ID: Hi, This is Dr Bhawana Gupta from India. I want to do transfection in floating cells either in U937 or in HL60. I procured Fugene hd from Roche. Do you have any experience of tranfecting floating cells and making stable cell lines. I couldn't get any specific protocol regarding selection of these tranfected floating cells and how to separate live cells from dead once, because we can not spin such a lower no of cells during selection. If you have related knowledge, please share with me. I am in acute need. Thanks & Best Regards Dr. Bhawana Gupta M.V.Sc. DISCLAIMER: The information in this email is confidential and may be legally privileged. It is intended solely for the addressee. Access to this email by anyone else is unauthorized. 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From sticher from bioc.uzh.ch Fri Aug 14 09:33:51 2009 From: sticher from bioc.uzh.ch (Patrick Sticher) Date: Fri Aug 14 10:49:33 2009 Subject: [Cell-biology] Practical Course in Biomolecular Modelling Message-ID: <4A8575CF.2050306@bioc.uzh.ch> Dear colleagues, please be informed that online applications are accepted for the following course: 8TH NCCR PRACTICAL COURSE IN BIOMOLECULAR MODELLING January 10 - 15, 2010 Kandersteg, Switzerland http://www.structuralbiology.uzh.ch/course2010.asp Course topics include Simulation techniques, force-field development, conformational search, computation of free energy and entropy, treatment of electrostatic forces, simulation of folding, comparison of simulation with experiment This course is primarily directed to PhD students and postdocs from experimental structural biology groups wishing to learn more on biomolecular modelling. The course format will include morning lectures and late-afternoon/early evening tutorials, and provide ample opportunities for discussions with experts and fellow participants. Participants will be invited to bring own problems for tutorials and/or discussion. The course will be organized as a winter retreat in the Swiss Alps offering a stimulating learning atmosphere with the afternoons available for informal participation in discussions, reading and self-study or recreational activities in the area. Interested candidates are encouraged to apply online on http://www.structuralbiology.uzh.ch/course2010_application.asp. Application deadline will be October 16, 2010. We will be able to accept 20 participants to this course. Best regards, Patrick Sticher -- _________________________________ Visit the NCCR on the Internet www.structuralbiology.uzh.ch Dr. Patrick Sticher Moser NCCR Scientific Officer Institute of Biochemistry University of Z?rich Winterthurerstrasse 190 CH - 8057 Z?rich Phone +41 / (0)44 / 635 54 84 Fax +41 / (0)44 / 635 59 08 Mail sticher@bioc.uzh.ch From kevcarby from gmail.com Tue Aug 18 13:37:43 2009 From: kevcarby from gmail.com (Kevin Carbajal) Date: Tue Aug 18 18:00:56 2009 Subject: [Cell-biology] Tips for Retrovial Transfection Message-ID: <8bc739800908181137m411c9353gfe9a17feaee904@mail.gmail.com> I'm not getting any positive cells in my titers. I'm using 293T, lipofectamine, and a Nature protocol with the DNA described therin (a GFP reporter). (Paper: Retrovirus-mediated single-cell gene knockdout technique in adult newborn neurons in vivo, Tashiro and Gage). Pretty standard protocol as far as I know. I didn't purify the virus by ultracentrifugation and just did titers with filtered viral sups instead. No positives by FACS. Where are the most common pitfalls? As far as I can tell, my cells are good, DNA is good, Lipofectamine is expensive, and the protocol was followed with a small change. Thank you for any help! -- From Jan.Visser from rug.nl Thu Aug 20 09:41:47 2009 From: Jan.Visser from rug.nl (Jan Visser) Date: Thu Aug 20 11:38:45 2009 Subject: [Cell-biology] Where to buy pentosidine? Message-ID: <4DFD67B25B5343ABA9873BC1D0085709@192168138> Hy Mr.Dunn, I saw in Google your message from 1996 about "Where to buy pentosidine?". I have the same question. Did you got an answer and was there any company who delivered this compound? Kind regards, Ing.Jan Visser University Centre for Pharmacy Department of Pharmacokinetics, Toxicology & Targeting Ant.Deusinglaan 1 9713 AV Groningen The Netherlands tel. 31-50-3633280 fax 31-50-3633247 e-mail: Jan.Visser@rug.nl From d_loeb from gmx.de Fri Aug 21 08:21:22 2009 From: d_loeb from gmx.de (Daniel Loeb) Date: Fri Aug 21 10:56:49 2009 Subject: [Cell-biology] Trypsin Message-ID: <20090821132122.136010@gmx.net> Hi, I have a problem with detaching my cells: I use 1x Trypsin (0.5 mg/ml) with EDTA to detach HeLa cells. But after incubation for some minutes at 37 ?C there are huge cell aggregates that do not seperate, even when I try to resuspend them by pipetting up and down. I tried it hard, but no chance. Moreover, when I want to pellet the cells, the aggregates are not soluble and because of that, I'm not able to centrifuge them down. Besides that, the suspension of trypsin and cells shows a strange behaviour on the plate: when I try to take out the suspension from the plate with the pipet, a big amount of suspension tries to redistribute on the plate even if I hold the plate slantwise. Perhaps some strange effects of surface tension. I hope somebody can help me. Perhaps its a simple mistake I am doing, because I am relatively new in that cell culture things. My colleagues say a little bit of that 'redistribution effect' is normal. Cheers, Daniel -- GRATIS f?r alle GMX-Mitglieder: Die maxdome Movie-FLAT! Jetzt freischalten unter http://portal.gmx.net/de/go/maxdome01 From sfanz from rediffmail.com Fri Aug 21 00:39:28 2009 From: sfanz from rediffmail.com (Aftab Hussain) Date: Fri Aug 21 16:52:57 2009 Subject: [Cell-biology] (no subject) Message-ID: <20090821053928.31385.qmail@f4mail-235-239.rediffmail.com> Can anbody please clarify the following doubts I have? Phytases are enzymes usefull in hydrolzing phytate ie.the organic source of phosphorus. Acid phosphatases are enzymes which solubilize inorganic phosphorus. In the literatue available it is mentioned that phytases are a gruop of phosphatases and some people say phosphatases are a group of phytases Which is correct? and I would like to know the classification of both phytases and phosphatases. could anbody in this field help me out Aftab from Bangalore India From mjalali from stedwards.edu Thu Aug 27 23:52:14 2009 From: mjalali from stedwards.edu (Mina M Jalali) Date: Fri Aug 28 00:43:18 2009 Subject: [Cell-biology] Protein Concentration Question Message-ID: <16116668.356561251435134219.JavaMail.root@mail2.stedwards.edu> Hey Erik, I saw your question posted on bio.net and was wondering if you ever figured out the answer. I had a similiar question: What is the concentration of a protein in a cell in molarity? I know that proteins account for 15% of a cell's overall concentration and that the molar mass of a protein is 50,000 g/mol. If you could help me figure out the answer, that would be great. Thank you! Mina Hi netters. Can anyone be of help by telling me where/how I might be able to find/calculate a "typical" concentration (mg/ml or something of the like) of a housekeeping protein like tubulin in a "typical" mammalian cell. I need a ball park figure. As far as I know, about 16-20% of a cell's weight is protein (but what's the typical weight of a "typical" mammalian cell?), volume is readily estimated from cell diameter, and a highly expressed protein could be about 1% of the total protein in a cell (so for, say tubulin, about 0.1-0.2% of the cell is protein by weight (?)). Any suggestions, info???? Please email. Cheers, erik