Pierce Super Signal
S K Marquardt
smarquardt at EARTHLINK.NET
Fri Oct 10 13:16:22 EST 1997
At 03:44 PM 10/10/97 GMT, Dima Klenchin wrote:
>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
>: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...
Doesn't that sound a little like polyvinyl alcohol?
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