Pierce Super Signal

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Fri Oct 10 10:44:56 EST 1997


In article <7opiGDf98QgB-pn2-tDOYuITQR.kt at RNAWORLD>, PGegen at UKans.edu wrote:
:On Sun, 5 Oct 1997 06:21:28, Juha Saarikettu 
:<juha.saarikettu at panther.cmb.umu wrote:
:
:> Does anybody have experience about Pierce Super Signal 
:> labeling/detection for southerns. How does it compare in sensitivity to 
:> DIG-system and radioactive labeling/detection. 
:
:I'd like some first-hand information, too! Our lab has tried the 
:SuerSignal kit but gotten terribly high background--virtually the 
:entire film was black after a 30-sec exposure. I suspect that we 
:used way too much antibody;  our postdoc probably used dilutions 
:recommended for colorometric detection because the instructions with
:the kit contradicted the instructions with the antibody itself... We
:haven't gone back to try any variations, and were wondering whether 
:to get a refund or try to optimize the signal. 

I poste similar Q a year ago and from may replies a clear answer has 
emerged: Pierce's SuperSignal is far superior to competitors. So we 
bought it and it works almost as advertised PROVIDED THAT IT 
WAS OPTIMIZED (something that has to be done always and with everything 
anyway). 

:I should say that since the blocking reagent that Pierce supplies is
:proprietary, we have no idea how to optiize blocking, yet Pierce 
:says this is a ccritical step in getting good images!

They want to sell more of their stuff. We use 5% milk and everything is
just fine. Of course, if primary ab are crap, nothing will ever help 
(we've some of that too).


:have experience with Pierce's blocker, or with using a normal 
:BSA-based blocker with the Super Signal kit?

I do. It does work in 15 min similarly to ~ 1hr with milk and it does 
kill most of the signal in 1 hr - something milk never does. We 
tried it also in immunochemistry - it decreased background AND 
signal down to zero. Wonder  this stuff is...

Dima



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