western blotting

Dima Klenchin klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Fri Mar 23 18:00:00 EST 2001


caosongsong at yahoo.com (songsong cao) wrote:
:I am doing western blotting now. The primary antibody
:I got is serum and I have no idea how many times I
:should dilute it before use. Could you give me some
:suggestions? Thank you.

There is only one suggestion that can be made:

Run many identical lanes of you protein sample
and do western with each of them separately using
different dilutions. Then pick the one that gives lowest
backgrounds while still providing sufficient sensitivity.
The range 1:100 to 1:10,000 with increrment of dilutions
of 1:5 or 1:10 should cover just about any possible
combinations of antigen and antiserum.

Sorry, there really is no other way of you intend to
do it right.

        - Dima




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