From: Ronald Rabin (rr84g@nih.gov)
Date: Tue Mar 18 2003 - 17:15:12 EST
We are commencing on experiments that will require human PBMC to be frozen prior to stimulation and analysis. Some cell surface markers (CD62L, CCR7) are lost in the process so that staining for these markers after thawing is useless. We could stain prior to freeze down with the tyramide system that would "deposit" FITC into the membrane of the cell; Presumably the CD62L+ cells would remain FITC+ even when CD62L itself is shed. However, scaling an enzyme linked system up to large number of cells seems tricky and expensive, so I did not pursue this approach. Is there any other way to do this? Perhaps a tag that deposits onto the cell after exposure to light that could be conjugated to an antibody or purchased as an avidin conjugate? ron > Ronald L. Rabin, M.D. > Senior Staff Fellow > Laboratory of Immunobiochemistry > DBPAP/OVRR > Center for Biologics Evaluation and Research > U.S. Food and Drug Administration > 29 Lincoln Drive (MSC-4555) > Building 29, Room 129 > Bethesda, MD 20892-4555 > > phone: 301.496.8806 > fax: 301.402.5177 > email: rr84g@nih.gov
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