blocking for western blot

Jim Kami jakami at ucdavis.edu
Mon Apr 24 12:21:20 EST 2000


I've used 5% non-fat dry milk for years with no problem for both
westerns and ELISA. The only "trick" is to dissolve the milk in water
first and then add 10X PBS. The milk seems to dissolve faster and more
completely without salt present.


Jim Kami
Department of Agronomy
Hunt Hall Rm 272
University of California
1 Shields Avenue
Davis, CA 95616-8515 USA
email: jakami at ucdavis.edu
Tel: (530) 752-9982
Fax: (530) 752-4361

"Why does Common Sense always seem to be the least common sense ?!?"


soenke behrends wrote:

> 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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