detection of phosphorylation

Ruth Knuechel-Clarke t866625 (Ruth.Knuechel-Clarke@klinik.uni-regensburg.de)
Wed, 25 Jan 1995 20:52:35 +0100

To all,
we are working on human cancer cells (bladder cancer), and have
defined the number and regulation of the Epidermal Growth Factor
receptor via flow cytometry and ligand binding assays. We would like
to relate the findings to the degree of activation of the EGFR and
have done so in Western blots. However all attempts to do the latter,
i.e. staining of the phosphorylated site of the receptor for flow
cytometry gives unreliable and varying results. Antibodies Py20
and others against phosphotyrosin have been used, and various
protocols of fixation have been tested systematically in the last 6
months. We would be very thankful for comments of people who have
experience with the use of antiphosphotyrosine antibodies or
related portocols in flow cytometry

Ruth Knuechel, MD
Senior Pathologist
University of Regensburg
FRG


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