At 15:15 06/05/02 -0500, you wrote: > >Greetings, > >Can anyone tell us if (IgG) anti glycophorin A antibody will cause >substantial changes in light scatter (particularly forward scatter) of >(human) RBC? We seem to see decreased forward scatter to the point of many >of the RBC almost being off scale to the left, and increasing the FSC gain >doesn't really help to bring these back on scale. This was just observed >today and the investigators will have to go back and look at the cells >microscopically after staining but I thought I would ask the group. > >Should they leave out added Ca and Mg in their binding and wash buffer, >i.e., are these (Ca/Mg)more of a problem with antibody staining of RBC than >lymphocytes? > >Thanks for any information. > >Ray Hester >College of Medicine >Univ. of South Alabama >rhester@jaguar1.usouthal.edu > > > Hello Ray, Ive been looking at RBCs and RBC microvesicles by flow, I generaly just add 5ul of anti GPA to 50ul whole blood and incubate for 20mins on ice, dilute 2000-fold in PBS/2%Fcs (+Ca/Mg) and put them through the flow. I can't say that i've noticed any problems: I use DAKO PE GPA Best regards, Arnold _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/ Arnold Richard Pizzey Department of Haematology Royal Free and University College London Medical School 98 Chenies Mews London WC1E 6HX U.K voice: +44 020-7679-6234 Fax: +44 020-7679-6222 email: a.pizzey@ucl.ac.uk _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/
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