Human B cell staining
Brian Rabinovich (brian.rabinovich@utoronto.ca)
Wed, 3 Jul 1996 02:47:16 -0400
I've been using Pharmingen FITC labelled anti-human CD19 mAbs to label B
cells from human peripheral blood, as part of a two colour assay assessing
B7.1 and B7.2 (both labelled with PE) levels on B cells. I am staining 75
microliters of whole EDTA anticoagulated blood with 15 microlilters of
antibody. I am unhappy with the sharpness, FL1 intensity, and % gated of
single positive cells (i.e FITC-CD19 only) I am obtaining. When gating on
lymphocytes = G1 on a SSC v.s. FSC plot the CD19 positive cells (i.e. LR
quadrant of FL2 v.s. FL1 dot plot of G1) from a FITC-CD19 only sample on
average make up only 7-8% of gated cells. What might be a better marker for
B cells? CD22?
CD ROM Vol 2 was produced by 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:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
