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I have recently discovered this site. I have a question regarding=20
reagents. We have been testing the Caltag monoclonal kappa/lambda/cd19=20
antbody cocktail and comparing it to the same BD combination to try and=20
get a brighter CD19 signal for use in a clinical flow lab. I have=20
noticed a couple of cases in which the BD reagent shows no evidence of a =
light chain restriction but the Caltag reagent does (usually a very low=20
percentage, dim kappa). Has anyone else seen this? The population=20
could not be confirmed with polyclonal kappa and lambda from BD or Tago.
Eric,
We have been working through some of the issues about kappa/lambda =
staining over the last 12 months. We have found (and been told by others =
- principally Carlton Stewart) that choice of flourochrome is important. =
CD19 density on mature B cells is quite low so if you choose to use CD19 =
to select out your B cells you need the brightest flourochrome available =
to get good signal - CD19PE is the only way to go in this regard (thanks =
Carl !).
An excellent alternative to CD19 is to use CD20 - higher antigen density =
(also expressed on a small subset of peripheral T cells but at a low =
levels so this is never a problem in our hands). We now use a cocktail =
of polyclonal kappa or lambda FITC/CD20 PE/CD45 PerCP (all BD reagents) =
and are extremely happy with the results.
We haven't evaluated the Caltag reagent so I cannot make a direct =
comparison.
Hope this helps :-)
Peter Chapple
Peter MacCallum Cancer Institute
Mebourne AUSTRALIA
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web