questions
Meenakshi Roy (meena@macsun.stanford.edu)
Mon, 10 Jun 1996 11:02:27 -0700
Hi everyone,
I have several questions about apoptosis assays and a question about B
cell subsets:
1. I have recently started using 7-aminoactinomycin D (without
permeabilisation) to detect live,apoptotic and dead cells. Is this a
popular/reliable method? I have heard that annexin-5 is a reliable method.
Has anyone had any problems with this?
I am basically looking for a method which will allow me to phenotype the
apoptotic cells by using FITC and PE dyes at the same time as assessing
apoptosis.
Also, is there a permeabilisation method which I can use to both detect
cells in G1,S,etc phase and also phenotype these cells on the FACS. (7-AAD,
annexin?)
2. This is a B cell question. What percentage of IgG and IgA positive B
cells can you detect in the mouse spleen, and lymph nodes? Do you need to
first gate on a B220 or CD19 positive population? What are good antibodies
to use for this purpose?
I would really appreciate any advice,
Dr. Meenakshi Roy
Dept. of Pathology, Stanford University
Stanford University/Foothill research center
R205, 4005 Miranda Avenue
Palo Alto, California, CA 94304
FAX: 415-858-3986
email: meena@macsun.stanford.edu
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