From: Luis Felipe Flores-Suarez (suarez@rummelplatz.uni-mannheim.de)
Date: Thu Apr 23 1998 - 09:03:28 EST
Hallo! I hope I can find some help. I am looking for the expression and detection of tissue factor antigen on the surface of HL-60 cells, and so far I have not had the results I thought. I seem to find few expression. The cells I have worked with are between the 15 and 25th passage, and usually the expression is looked for 1 or 2 days after passaging them. I use a murine monoclonal anti-human anti-tissue factor antibody (from American Diagnostica Inc) and then I detect it using a rabbit-antimouse FITC or PE conjugated antibody. Can someone give me data about the amount of antigen molecules present on the surface of the cells? Is this quantity overinduced by the addition of lipopolysaccharide (specifically from E. coli O:55 B:5)? If that occurs significantly, how many hours of incubation with LPS are needed to obtain the full-blown, maximal expression of tissue factor antigen on the surface of HL-60 cells? I hope someone can give me a hand on the item. Thank you very much in advance. Luis Felipe Flores-Suarez, M.D. Mannheim, Germany.
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