Dear Melanie Caltag's Fix & Perm is a stand alone system. If you have already done any previous lysing, you cannot use Fix & Perm as a an additional step. This will be clear in the scatter pattern which will not be normal. Regards Bob Johnson Caltag "O'Donahue, Melanie ." wrote: > Charles, > I have begun doing cytoplasmic staining on CLL patients that do not exhibit > surface light chains. I use the stained and fixed tube, assuming there is > enough left after acquisition. I add 100 ul reagent B from Caltag's fix and > perm and more light chain antibody and incubate for 15 minutes. Then I wash > in PBS, resuspend, and acquire again. Quite often the light chains appear! > Thanks for the CD 38 reference. I'll definitely read that. > Melanie O'Donahue > Mayo Clinic Scottsdale > > -----Original Message----- > From: Repetti, Charles [mailto:crepetti@aml.com] > Sent: Monday, December 11, 2000 12:59 PM > To: cyto-inbox > Subject: RE: Several Questions > > Melanie; > The CD38 references are in Blood, Sept 15, 1999Vol 94,pp1837-1839; > pp1840-1847 (the flow CD38); pp 1848-1854. CD5-FITC, CD38-PE, CD19-PE:CY5. > The CD38 is usually discreet, almost all negative or mostly positive in the > 2nd or 3rd decade. > > CLLs without K,L: have very little experience with intracellular, would > recommend intsead having alternative k,l clones for surface staining. Would > be interested in any info on cytoplasmic stains. > > good luck > Charles > > -----Original Message----- > From: O'Donahue, Melanie . [mailto:ODonahue.Melanie@MAYO.EDU] > Sent: Friday, December 08, 2000 4:36 PM > To: cyto-inbox > Subject: Several Questions > > Hello all, > There are several things I'm trying to troubleshoot and/or improve and I > would appreciate any input from my fellow flowers. > Are any of you doing platelet hyper aggregation studies with flow? > Also, have you had any experience running CD 38 on CLL patients? It's > supposed to be a prognostic indicator. One of our heme onc docs is > interested in it. > On CLL patients that don't express light chains, do you routinely run them > intracellularly? What rate of success do you have with that? > Final question. Do you run your light chains in two separate tubes? Is their > a reason for that? We are having problems with our three color > (kappa-lambda-CD 19) tube and I'm wondering if I should be running two color > instead. > Thanks for any input. Have a great weekend! > Melanie O'Donahue, MT, ASCP > Mayo Clinic Scottsdale
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