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).
CD13 is expressed on the surface of early committed progenitors and mature granulocytes and monocytes (GM-CFU), but not on lymphocytes, platelets or erythrocytes. It is also expressed on endothelial cells, epithelial cells, bone marrow stroma cells, and osteoclasts, as well as a small proportion of LGL lymphocytes. CD13 acts as a receptor for specific strains of RNA viruses and plays an important function in the interaction between human cytomegalovirus (CMV) and its target cells.