Amersham Hybond N problem

Paul N Hengen pnh at cockleberry.ncifcrf.gov
Fri Mar 1 13:09:59 EST 1996


BRITTA SCHULZ (schulzb at pz350.ipsp.uni-hohenheim.de) wrote:

: Who can help me to find out, which company produces the uncharged membrane
: formerly sold by Amersham as Hybond N ?

: The problem is , they are now selling a new membrane, calling it Hybond N as 
: well, but it is very different. However, since we have the new Hybond N all 
: we get is black autoradiographs (we are using the Dig-system) The new 
: membrane is very thick and stiff, it doesn't look like nylon 
: at all, and it is produced by another manufacturer. Since I would like to go 
: on with my work without too much delay and the Amersham people did not want 
: to tell me their former supplier, maybe someone here knows something.

I welcome you as a new member of the Great Membrane SWAP Club! You wrote that
you are using a DIG system, but didn't say what the detection chemistry is.
After MUCH trouble with the membrane supplied with the Tropix Southern Light
kit, I've settled on a new membrane. You should also know that the companies
are STILL fighting over patent rights and their membranes might change in the
future, so be careful out there! I recently got a letter from a company who
SWAPped membranes on me stating that their membranes are the same as before
(refuting a claim made by a competing company), but don't believe it! I have
the blots to prove otherwise.  Anyway, although I did not test the membrane
you are using and I am using a biotin based system, here are my results of
membrane testing if you would like to shop around...

-------------------------------------------------------------------------------
-------------------------------------------------------------------------------
Last year I posted a message warning netters about a membrane SWAP done by
Tropix with their Tropix Chemiluminescent detection kit called Southern Light.
For more information about what happened and the response of Tropix, see the
article below or you can search the archives for my posting to find out what
the problem was.

@article{Hengen1995Dectibs,
author = "P. N. Hengen",
title = "Methods and reagents - Floaties in the gene pool",
journal = "Trends in Biochemical Sciences",
volume = "20",
number = "12",
pages = "522-523",
month = "dec",
year = "1995"}

After doing MANY side-by-side comparisons of different membranes using the
same kit and my proven protocol of semi-dry transfer. The results follow:

Membrane                               Result
--------                               ------
"Old" Tropilon Plus Membrane           **** Excellent Membrane (NLA)
"Old" MSI MagnaCharge                  **** Excellent Membrane (NLA)
Pall BioDyne A                          *** Pretty Good        (1st choice) 
S&S Nytran Plus                          ** Some spotty bands  (2nd choice)
BioRad Zeta Probe GT                     ** Spotty bands
BioRad Zeta Probe                         * Total loss of some bands
Pall BioDyne B                            * Weak signal, no background
"New" MSI MagnaCharge                     - Dark Background Problem
Pall BioDyne Plus                         - No Bands, Not Acceptable
"New" Tropilon Plus Membrane              - No Bands, Not Acceptable


**** = Excellent
***  = Good
**   = Okay
*    = Poor
-    = Don't waste your time
NLA  = No Longer Available
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Oh, BTW, I hate what the new WWW hyper-mail thingy does with spaces!!! >:-{
If you want a readable version of this table, I'll send a copy by e-mail.

*******************************************************************************
Paul N. Hengen, Ph.D. (pnh at ncifcrf.gov)
National Cancer Institute--FCRDC--Frederick, Maryland 21702-1201 USA
phone:(301) 846-5581 fax:(301) 846-5598
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