Removal of antibodies in Co-IP

Simon T. Buttgereit tbutt at web.de
Fri Jun 4 03:20:34 EST 2004


Dr Engelbert Buxbaum wrote:

> Either crosslink the antibody to a carrier (e.g. agarose beads) or use
> denaturing isoelectric focussing instead of SDS-PAGE for separation. 

Have tried ist, but activity of antibody went down dramatically.

> What kind of detection are you using? 

I just want to cut out differential protein bands between two induction 
timepoints for MALDITOF. 2D-PAAG should be performed later.

With metabolically labelled 35S or
> with 125I labelled proteins the presence of the non-radioactive Ig chain
> would not interfere with detection. 

thats right, but they MALDI-Guys don´t like radioactivity...

I have found some kits for antibody removal. Have an idea how they work?

Thanx
Thomas




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