From: biocytex@biocytex.com
Date: Mon Jun 29 1998 - 11:53:51 EST
Dear Jan, Whatever the instrument I use, I would avoid placing the histogram of isotype control close to the first channel. This induces over-evaluation of its "Mean" value. In order to measure low levels of expression such as cytokine receptors, I would try to place the control between the 1st and the 2nd decades i.e having a Mean value of 10. This leaves plenty of space on the right for positive stainings and log amps most often show good linear response in the middle range. Also I would avoid murine IgG2a MABs since these show rather high non specific background with HL60 due to Fc receptor binding. Prefer IgG1 MAbs if available. Hope this helps. P.PONCELET R&D Director BioCytex
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