Peptide phage library screening by Flow

From: Raj Nathan (rr5845@hotmail.com)
Date: Fri Mar 31 2000 - 11:49:15 EST


Hi
We use the elisa technique succesfully in our laboratories to detect phage
bound to the leukemic cells. We use the monoclonal mouse anti-M13 antibody
conjugated with HRP to detect the bound phage. But our repeated attepts to
detect bound phage by flow has failed. I have used Mouse anti-M13
unconjugated primary antibody followed by using a anti mouse goat secondary
which is conjugated to FITC. A couple of papers have shown bound phage
detection by using a sheep anti-M13 antibody followed by secoundary
conjugated with FITC with success, But the company Parmacia stopped
production of this antibody. The company has not tried out the mouse anti
M13 primary to check phage bound to cell by flow. So i would be very
grateful if anybody could give me information on protocols tried out or to
use or information of antibodies that may be used for detection of the bound
phage by flow. I have also tried out anti HRP antibody against the anti-M13
HRP primary that we use for the ELISA but still could not show the detection
of phage bound to the cells.
Thanks
Raj
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