This question is posed on behalf of the person below. Please reply directly to this person, or to the list with yoru comments. Do not use the REPLY function - it will not go anywhere! Reply to empig@genome2.hsc.usc.edu ------------------------------------------------------------------- Posted message: >My question is regarding virus binding to HeLa cells. Briefly, we mix >virus supernatant with cells for 2h @ 4oC, then come in with a virus- >specific 1o Ab, followed by a FITC-conjugated 2oAb then FACS analysis >to differentiate between cells that bound virus from those who didn't. >My problem is that with HeLa cells, I get a second peak on my >histograms. People have suggested that perhaps my binding assay damages >the HeLa cell membrane and allows non-specific incorporation of FITC, >thus resulting in a peak on the right of the histogram. Does anybody >have experience in looking at virus binding to HeLa cells and has gotten >around this problem with possible HeLa cell membrane damage? Also, has >anybody done LDLR cell surface expression on HeLa cells using FACS? >Thanks for your help. > >Cyril Empig > >My email is: empig@genome2.hsc.usc.edu > ------------------------------------------------------------------------ This message posed by JPR for the above person. J.Paul Robinson, Ph.D. Professor of Immunopharmacology Director, Purdue University Cytometry Laboratories EMAIL: robinson@flowcyt.cyto.purdue.edu WEB http://www.cyto.purdue.edu Phone: (765) 494 0757 FAX (765) 494 0517
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