Expression of HLA-B27 in peripheral blood leukocytes
Expression of HLA-B27 in peripheral blood leukocytes
This experiment is based on "Color Atlas of Practical Flow Cytometry", edited by Shukui Wang and Zhenying Zhou.
Operation method
Expression of HLA-B27 in peripheral blood leukocytes
Materials and Instruments
CD3 Monoclonal antibody HLA-B27 Monoclonal antibody Whole blood Move 1. Take 30 μl of fluorescein PE-labeled CD3 monoclonal antibody and FlTC-labeled HLA-B27 monoclonal antibody, and 50 μl of EDTAK3-anticoagulated whole blood, and then mix well and stain for 15 min (or 20 min) at room temperature. For more product details, please visit Aladdin Scientific website.
Hemolytic Agents Paraformaldehyde
Flow Cytometry
2. Add 2 ml of hemolytic agent to dissolve the red blood cells (10 min).
3. Remove cell debris by centrifugation at 1500 r/min, and fix with 1% paraformaldehyde (15 min).
4. Measurement on the machine. After measuring the standard microspheres and adjusting the Fl1 voltage, do not change the FL1 setting. Set the "gate" by CD3-PE/SSC to circle the CD3-positive cells (T lymphocytes), and detect the average fluorescence intensity of their B27. A specimen is negative if its fluorescence intensity is less than the standard microsphere fluorescence intensity, and positive if the opposite is true.
5. Give the flow cytometry results and issue a lab report.