blocking for western blot

flow deszo at uiuc.edu
Mon Apr 24 10:19:50 EST 2000


    I have used 2-5%BSA in TBST 0.5% Tween20 with good results.  Also I have
noticed that with the Pierce kit I had to lower the secondary concentration
to reduce the background.

Eric
"soenke behrends" <behrends at plexus.uke.uni-hamburg.de> wrote in message
news:8dmilq$e4e$1 at rzsun03.rrz.uni-hamburg.de...
>
> Dear netters,
>
> we are routinely using milk powder as
> blocking agent for western blots using
> nitrocellulose blots. However,
> we have noticed that when we leave out
> milk powder altogether there is no difference.
>
> We now want to try products like super signal
> from pierce where the company argues that you
> get stronger and better signals as compared to
> regular ECL solution. After our first experiments
> we see that background really becomes a problem
> with this solution and that we need a good
> blocking agent.
>
> Since we could leave out the milk powder in our
> protocol without any problems, we start to question
> whether it has any effect at all. Pierce offers
> different kinds of blocking agents, but they are
> expensive and available only in large quantities.
>
> We would be glad to hear about any tricks or hints
> and whether the commercial solutions compare favorably
> with 'home-made'-recipees.
>
> Thanks for your time and help
> Soenke
>
>
>






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