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We have successfully used competitive binding experiments to measure the binding
of HIV gp120 to the CD4 receptor. In this assay serial dilution's of gp120 are
added to cells, washed and stained with either Leu-3a (blocked by gp120) or
OKT4 (not blocked by gp120). The channel locations are calibrated for the
antibody binding capacity so that we could determine quantitative blocking
efficiency. This was also done using whole virus with good success.
Stephen P. Perfetto
Department of HIV Disease Prevention
Walter Reed Army Institute of Research
1600 East Gude Drive
Rockville, MD. 20850
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Subject: Labeled Virus
From: Dennis_Young@cis.ucsd.edu at Internet_Gateway
Date: 2/7/96 10:30 AM
Is there a better way to follow virus binding than R-18 "dequenching"
(Leary,JF, Notter,MFD, Kinetics of Virus Adsorption to Single Cells Using
Fluorescent Membrane Probes and Multiparameter Flow Cytometry, Cell Biophysics,
4, 63-76, 1982.)?
Our first attempt failed to see any rhodamine fluorescence. What would be a
good positive control?
FACSTAR+, 514 nm excitation, 560 dichroic and 600 SHORTPASS in FL-2.
Dennis
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* Dennis J. Young Voice : (619) 822-0407 *
* Flow Cytometry Core Facility FAX : (619) 822-0412 *
* University of California, San Diego USA e-mail: djyoung@ucsd.edu *
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