CD23 staining of B cells

From: Calman Prussin (CPRUSSIN@niaid.nih.gov)
Date: Fri Jun 21 2002 - 11:52:58 EST


Dear flowers,

I am providing analysis of clinical research samples in which the
investigators are following FceRI and CD23 (low affinity IgE receptor)
expression during a trial of immunomodulatory therapy. In order to process
the 120 samples efficiently, the cells were ficolled, fixed in 4% PFA,
aliquotted and cryopreserved at -80. We find this procedure works very well
for intracellular staining.

I have no experience staining for CD23 and would like to get some feedback
on what intensity of staining I should expect to see on B cells. We ran 6
allergic asthmatic donors, staining for CD19 PE and CD23 APC (both Caltag)
using saturating concentrations of mAb.

I am hoping to detect down modulation of the CD23 signal. However, if the
CD23 staining is already suboptimal, it will be more difficult to detect a
decrease in MFI.

I have placed the plots on the following web page:
http://tango01.cit.nih.gov/sig/cytokine/CD23.htm

Please comment on:

1. Relative brightness of this staining to what you are used to.

2. If your CD23 staining is significantly brighter, what clone (company) are
you using?

Thanks,

Calman
> _______________________
> Calman Prussin
> Laboratory of Allergic Diseases
> NIAID/ National Institutes of Health
> Bethesda, MD	20892-1881
>



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