Protein Expression Screening

Jayne Williams williams at sb.fsu.edu
Tue Jul 2 07:52:49 EST 1996


Hello Everyone,

I am a beginner and there is probably a simple answer to my question,
yet I have been unsuccessful in finding anything in the literature on
this:

I have transfected a plasmid (pMAMneo) into CHO cells (Lec-1 line), and
will soon induce expression of my protein (Thy-1: a cell surface
glycoprotein, 111 aa). I have already verified the presence of the gene
by PCR.

I'd like to quickly screen for protein expression by comparing SDS-PAGE
of cell lysates (induced vs noninduced). My question: how much lysate
(very roughly, how many cells) must I load to see my protein on the gel?
(Of course, I have no idea at this point what level of expression I will
get, so am only looking for a reasonable starting point.)

Any suggestions/references very much appreciated. (E-mail replies
equally appreciated).

Jayne Williams
Institute of Molecular Biophysics
Williams at sb.fsu.edu



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