Optimal working dilution should be determined by the investigator.
Assay Procedure
Take 100 μL peripheral blood anticoagulated by EDTA and add to the bottom of 5 mLtube,
Add appropriate amount of antibody to the bottom of flow tube mixing with the whole blood, incubate for 30 minutes at room temperature,
Add 2 ml1×RBC lysis buffer, incubate for 10 minutes after mixing, dissolve red blood cells (recommended: RBC lysing Solution 10×,Cat.: FXP001),
Sample tube is set to 1000 rpm centrifugation for 5 minutes, discard the supernatant,
Add 2 mLPBS wash buffer to resuspend the cells, then 1000 rpm centrifugation for 5 minutes, discard the supernatant,
Add appropriate amount of fluorescent-labeled Streptavidin and incubate for 20 minutes away from light at room temperature.
Repeat step 5.
Add 0.5 mLPBS wash buffer to resuspend the cells and detect by flow cytometry (sample should be determined on the day on the machine and can also be added fixation overnight at 4 °C then measured).
B220 antibody, CD45 antibody, CD45R antibody, GP180 antibody, L-CA antibody, LCA antibody, LY5 antibody, T200 antibody, Lca antibody, RT7 antibody, protein tyrosine phosphatase, receptor type C antibody, protein tyrosine phosphatase, receptor type, C antibody, PTPRC antibody, Ptprc antibody
Background
This is clustered as CD45RA, and is expressed on naive/resting T cells and on medullart thymocytes. In comparison, CD45RO is expressed on memory/activated T cells and cortical thymocytes. CD45RA and CD45RO are useful for discriminating between naive and memory T cells in the study of the immune system.