From: DEBORAH SHUEY GROVE (dsg4@genesis.ait.psu.edu)
Date: Thu Aug 10 1995 - 08:34:28 EST
We are setting up studies to look at surface antigens on human PBL. We have experience with rat splenocytes and lymph node cells and would like advice on working with human PBL. 1) We are considering staining whole blood vs. separated.comments? Population differences? 2) We would like to screen a panel of antibodies in as few samples as possible by doing several markers in a single group. We can use FITC, PE, and PECy5. Are there some combinations of markers that are best to use? For example, TCRalphabeta, CD4, and CD8 may be good to run together to determine what percentages of T are CD4 and CD8 and if any are double-labelled. Also, NK with a T cell marker as a negative control? 3)Any advice on the best controls for human PBL? LCA? 4) Any advice on adhesion markers? We expect changes in circulating populations. 5) Data analysis? Are there published examples of histograms that we can ue for comparison? Thanks in advance. Deb Grove Dept BMB Penn State
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