summary:chemiluminscent westerns

braam at DINGO.RICE.EDU braam at DINGO.RICE.EDU
Tue Mar 30 05:50:19 EST 1993


Most people suggest using Amersham-ECL for chemiluminescent westerns. 
Note that all replies were positive; there were no "worst" kits 
suggested.

AMERSHAM-ECL:
1.	"I have not done any extensive comparisons of systems available for
chemiluminescent westerns, but I have been using the ECL system from
Amersham (
with horseradish peroxidase as the secondary antibody) with very clean
results.
  Of course, background will increase if the membrane is not properly
blocked or
if excessively high amounts of antibody are used (probably because it is
not
washed off well.)
Good luck, Estelle Hrabak"

2."	I never used the kit myself (I usually work with DNA, not protein) but
a lot
of my labmates have tried the Amersham chemiluminescence kit and loved it.
It is extremely easy, reliable, sensative, and quantitave (or at least they
tell me).  In general, I have been very happy with Amersham quality and 
service, and I have a lot of confidence in their stuff.
	Dr. Leonard N. Bloksberg
	PreissJ at clvax1.cl.msu.edu
	Dept. of Biochemistry
	Michigan State University"

3.	"I have used Amersham's ECL kit with success for quite a while.  It
appears to be quantitative across a wide range, probably limited by the
film you use.  
Kodak AR or S appears to work quite well, Amersham do their own film for
ECL 
but I've never felt the need to buy it!  I transfer onto Hybond-C, 
also from Amersham.  Standard disclaimers....
Richard
R. P. Grant    <><                     rpgrant at molbiol.ox.ac.uk
Sir William Dunn School of Pathology   Fax. +44 865 275556
University of Oxford, UK.              Tel. +44 865 275565"

4."	Our group is currently using Amersham's ECL Western blotting
kit and we are satisfied with it. We get nice quantitative
bands from westerns and the chemiluminescence reaction lasts
for minutes, giving ample time for multiple exposures (usually
3 seconds is enough!). You have to wash filters carefully
before adding the substrate, otherwise the background glow may
give you nice black films.
Hope this helps.
Cheers,
Tapani Ronni
National Public Health Institute
Mannerheimintie 166
00300 Helsinki
FINLAND"

TROPIX:
"I highly recommend "Western-Light" by Tropix, Inc, Bedford, MA
(800) 542-2369.  The sensitivity of the reagent allowed
me to detect fairly quantitatively a low level of expression
from the E. coli chromosome.  Combined with a PVDF membrane
(Immobilon from Millipore) it was super. However, I did not
use the secondary Ab that came with the kit as it gave a
cross-reacting band with my antibody.  So I used the AP
conjugated Ab from Promega that I had used with the color
reaction.
Needless to say I have no affiliation with the
above companies, except that the people at Tropix were
very helpful and I have a vested interest in keeping
them in business.
Pat Foster
Boston University School of Medicine
Boston, MA
pfoster at med-itvax1.bu.edu"

BOEHRINGER:
"Boehringer, without a doubt - but if you can conjugate your own Ab with
alkaline phosphatase, and buy AMPPD cheaper than from B-M (eg. from
Tropix, in New Bedford), then you can make your own kit for cheap.  "
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