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?