Dear flowers: We are trying to measure LFA1 expression in human peripheral blood leukocytes pre to post stress. Our purpose is to 1) measure changes in numbers of leukocyte subsets that are expressing LFA1 and 2) quantifying LFA1 on leukocytes. My questions are: 1) Would it be valid to stain cells with only CD11a antibody to detect LFA1 as opposed to doing a double staining for CD11a/CD18? 2) If we should do a double-staining, what implications it would have to measure CD11a & CD18 double positive, and CD11a & CD18 negative or the other way around? LFA1 is a heterodimeric adhesion molecule with alpha (CD11aL) and beta (CD18) chains, therefor it appear to be reasonable to stain for both CD11a/18, but I wanted to clarify. My take on the literature is that some studies reported that they measured CD11a to detect LFA1,some studies measured both (double positive), and some studies used both antibodies for CD11a and CD18 separately to examine functionality of each chain. My last question is regarding quantitation (antibody density) of LFA1. We consulted with BD and was told that samples need to be in separate tubes for CD11a and CD18 if we were to quantitate both since they only have the capability to quantitate PE-conjugate reagents. Does anybody have any experience on other ways of doing this? Any comments or input will be greatly appreciated. Thank you. Suzi Hong, Ph.D. Dept. of Psychiatry UCSD Medical Center 619.543.5832
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:22 EST