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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