Pre-absorbing Polyclonal Ab's

Eric at
Wed Nov 27 03:59:21 EST 2002

g9610820 at wrote:
> Hi all.  I have raised polyclonal Ab's to my protein of interest
> (which was purified using a C-terminal His-tag) in a rabbit, and I
> seem to have co-purified some of the E.coli protein, as I am getting
> lots of non-specifics on my WB's.  I have heard that you can
> pre-absorb your Ab's against the proteins from E.coli to remove the
> non-specific Ab's (ie the Ab's specific for E.coli proteins), but I
> am not sure how to go about this.  Does anyone have any protocols for
> me?  Thanks.

There could be many problems, e.g., either the °1 and/or °2 Ab
concentration is too high.  However, a good reference is...

Using Antibodies: A Laboratory Manual
Ed Harlow, David Lane
Cold Spring Harbor Laboratory
ISBN: 0879695447

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