The 4A104D2 monoclonal antibody reacts with human CD117, also known as c-Kit , Steel factor receptor and stem cell factor receptor. A member of the tyrosine kinase receptor family, this 145 kDa molecule is expressed by hematopoietic progenitor cell subsets and mast cells. The interaction of c-Kit and Steel factor promotes proliferation and differentiation of hematopoietic progenitor cells and mast cell differentiation. CD117 is also expressed by melanocytes and plays a role in signaling and activation of these cells.
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 10 μL labeled antibody to the bottom of flow tube mixing with the whole blood, incubate for 20 minutes at room temperature away from light,
Add 2 ml1×RBC lysis buffer, incubate for 10 minutes away from light 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, then1000 rpm centrifugation for 5 minutes, discard the supernatant,
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).