Calman- My sense is that the isotype control provides a "limit of sensitivity" that should be considered in the interpretation of samples with low antigen density. I agree with you that it makes less sense to subtract the isotype results as "background". Albert Albert D. Donnenberg, Ph.D. Interim Director Bone Marrow Transplant Program, UPCI Phone: (412) 624-9596 Fax: (412) 624-9624 -----Original Message----- From: Calman Prussin [mailto:CPRUSSIN@niaid.nih.gov] Sent: Thursday, January 06, 2000 11:43 AM To: cyto-inbox Subject: MESF and correcting for isotype matched controls We are quantitating cell associated antigens using standardized beads and then generating an MESF from the standard curve generated from the beads. The question has come up whether or not to "correct" the MESF values obtained from the specific antibody by subtracting the MESF values obtained for the isotype matched controls. The problem with correcting the values is that it has little effect on the samples with high density of Ag, but has a large effect on those with low expression. As such, if we run replicate samples on the low expressing samples we find a larger amount of variation in the corrected MESFs. My bias is not to use it, as it seems to introduce more "noise" into the system. Your thoughts? Thanks! > _______________________ > Calman Prussin > Laboratory of Allergic Diseases > NIAID/ National Institutes of Health >
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