Flow cytometry reagent needed
ladasky at my-deja.com
Thu Aug 29 16:40:58 EST 2002
I have just spent two fruitless hours searching the Internet and
various catalogs. I am looking for an anti-mouse Ig to which PE-Cy5
has been conjugated. I can't seem to find anyone who makes it. Why
Manufacturers may have reasoned that one would only choose to use
PE-Cy5 in a three-color flow cytometry experiment. They probably
think that, if you're doing three colors, at least two of them are
probably antibody labels. A second-step reagent would not easily be
confined to labeling your antibody of interest.
Still, anti-mouse Ig-PerCP is available, os maybe there is some other
subtle reason I have missed. I would rather not use PerCP, as some of
the antigens that I am examining are not very abundant.
In my case, I am using YFP as my first label, and I am trying to
perform a quantitative measurement of surface antibody staining using
another color. The changes are rather subtle, at most five-fold and
typically two-fold. My attempts to perform that quantitative
measurement using Cy3 or PE as the antibody labels have been
disappointing. I believe that the compensation circuitry is adding
too much noise and systematic error to my measurements.
I have had some success using Cy5 as the second color, on a flow
cytometer with two lasers. That flow cytometer is not especially
convenient for me to use. On the other hand, a single-laser flow
cytometer has just become available. I would like to try to move my
assay over to the single-laser machine.
John J. Ladasky Jr., Ph.D.
Department of Biology
Johns Hopkins University
Baltimore MD 21218
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