g9610820 at campus.ru.ac.za 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
1998
http://www.amazon.com/exec/obidos/tg/detail/-/0879695447//002-9330805-2829610?v=glance&s=books
