Background on blots

postmaster at ideaexplore.net postmaster at ideaexplore.net
Fri Nov 24 20:43:05 EST 2000


Hi,

By the way, you can attract more attention to your message by writing a Subject.

You should always block a western blot.  I assume you are already blocking at least 1
hour.  Overnight might help somewhat, but probably not much.  It's worth a try since it's
easy.

Also, consider the antibody you use.  Is it specific?  If it is monoclonal and not
specific, then you have to get another antibody.  If it is polyclonal,
affinity-purification may clean up the background problem.  That takes some time however.

I'm just speculating, but perhaps a quick way to reduce staining of extra bands is to mix
the antibody with a protein extract that doesn't have your target protein, but does have
some of the others, and then stain your blot.  That might bind up the non-specific
antibody.

First, try O/N blocking.  Good luck.

Larry Baum

Check out my website for more ideas:   IdeaExplore.net

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