Protocols to quantitate westernblots

[Jeff Rawlings]

In article <136157 at cup.portal.com>, Zekeman at cup.portal.com (Enrico Ami-tox
gilberti) wrote:

> Hello all, I am searching for methods that deal with quantitation of immuno-
> blots.  I have plenty of software but no real protocol.  All of the software
> I have alludes to measuring blots but non actually commits a logical protocol
> for it.  Any ideas, thought, or comments on this.  I figure the method will
> only be semi-quantitative considering all of the variables that are involved.
> Any help would be grateful.

I worked out a protocol using I-125 labeled secondary antibody, followed
by visualization with a phosphorimager. If you have a source of puified
antigen, of known concentration, you can use that as a standard on the
blot. I believe such a protocol can be quantitative.

Cheers,
--Jeff

-- 
Jeff Rawlings                                   Internet Consultant
rawlings at silcom.com                                             and
rawlings at cco.caltech.edu                         Research Biologist