---------- From: Eric Hsi[SMTP:ehsi@luc.edu] Sent: Wednesday, 1 January 1997 5:05 To: cyto-inbox Subject: kappa/lambda/CD19 reagents I have recently discovered this site. I have a question regarding reagents. We have been testing the Caltag monoclonal kappa/lambda/cd19 antbody cocktail and comparing it to the same BD combination to try and get a brighter CD19 signal for use in a clinical flow lab. I have 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 percentage, dim kappa). Has anyone else seen this? The population 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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