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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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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web