From: Albert Tai (acktai@exelixis.com)
Date: Wed Apr 09 2003 - 20:15:07 EST
Hi All, I have a couple elementary questions about 2 color compensation and hope someone with more experienced will be able to help me with my analysis. 1) I do compensations with FITC-conjugated (2nd)Ab to mouse primary Mab and PE-conjugated (2nd)Ab to Rabbit polyclonal Ab together with unstained cells. The XMean for FITC in FL-1H was 60 and XMean for PE in FL-2H was 270 after compensation. When present with my double-stained sample, the same cells used for compensation, the XMean on FITC reduced to 36 and XMean for PE reduced to 120. Is this observation reasonable ? 2) Also when staining cells with FITC-conjugated secondary Ab, there is roughly 5 folds different between unstained cells and cells only stained with FITC-conjugated secondary Ab, without primary Mab. I did not see that with cells stained with PE-conjugated secondary Ab alone which has same intensity as unstained cells. Any thoughts will be of tremendous help in designing my next experiment and thanks very much in advance. Albert Tai Signal Transduction Research Exelixis, Inc. P.O. Box 511 170 Harbor Way So San Francisco, CA 94083 (650)837-7228 (direct) (650)837-7240 (fax) e-mail: acktai@exelixis.com
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