Hi Calman, Based on what you have noted so far, I would be interested in reading whether you get the same variability upon subtraction of the MESF values for (unstained) controls. As to the subtraction of the matched isotype control values, it seems the "right thing to do" but others on the list may indicate better reasons not to. Cheers, Sam Sam Witherspoon sw11527@glaxowellcome.com Dept. of Receptor Biochemistry Tel. 919-483-3078 Glaxo Wellcome R&D Page 919-857-7768 5 Moore Dr. Fax 919-483-0585 RTP, NC 27709 > -----Original Message----- > From: Calman Prussin [SMTP:CPRUSSIN@niaid.nih.gov] > Sent: Thursday, January 06, 2000 11:43 AM > To: Cytometry Mailing List > 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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