seanpat at FMED2.UNCU.EDU.AR
Thu Nov 18 08:02:02 EST 1999
>Ok, let's say protein A binds to protein B which binds to protein C. If
>I pull down with an antibody to protein C (from cell lysates), should I
>expect to be able to pull down protein A with it and probe for protein A
>on a Western, or is it too much to hope for that everthing will stay
>together during the IP? I'm sure it depends on dissociation contants,
>but generally speaking...
A couple of thoughts.......
If your antibody requires denaturation to see the epitope you won't
IP anything - polyclonals which recognize multiple epitopes would be
better. It would also be unfortunate if your epitope was shielded in the
If you can also get antibodies to A and B (if their identity is
known) you should also be able to bring down the same complex using
different antibodies, a control that Harlow & Lane strongly recommends to
avoid embarrasing errors.
Try it and see, you never know.
Sean Patterson, Ph.D.
Catedra de Fisiologia, CC33
Facultad de Ciencias Medicas
Universidad Nacional de Cuyo
Tel: (0261) 420-5115 ext.2684/2747
Fax: (0261) 449-4117
e-mail: seanpat at fmed2.uncu.edu.ar
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