Patricia, We have been performing HLA B27 assays on the Coulter Xl for a couple of years now. Please refer to our paper"Utilization of commercial antisera and flow cytometry in HLA B27 typing" published in Cytometry 18,17, 1994. We use One Lambda HLA B27-FITC conjugated antisera and have found it to be monospecific. Our procedure is as follows: to 100uL of whole blood we add 2uL of the HLA B27 or isotypic control reagent. We incubate 15 minutes at RT in the dark. Wash once in PBS. Since we are using the QPREP system we add 100uL of fetal calf serum(as a protein stabilizer) and process on the QPREP. We then analyze on the flow cytometer. We find that this system works very well with a good distinction between positives and negatives. If we do not have a recent positive around to use as a control(we can use positive patients samples as controls for approximately two weeks without any preservation) we use CRISP cells provided by Phoenix Flow Systems. These cells are extremely stable and work well. Hope this is of some use to you. If you have any further questions do not hesitate to call or e-mail. Best regards, Ken Orr
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