4AE11 reacts with CR1 , the receptor for the complement component C3b /C4, composed of four different allotypes (160, 190, 220 and 150 kDa). CD35 antigen is expressed on erythrocytes, neutrophils, monocytes, B -lymphocytes and 10-15 % of T -lymphocytes. CD35 is catagorized as a regulator of complement avtivation. It binds complement components C3b and C4b, mediating phagocytosis by granulocytes and monocytes . Application: Removal and reduction of excessive amounts of complement fixing immune complexes in SLE and other auto-immune disorder.This clone is cross reactive with non-human primate.
Optimal working dilution should be determined by the investigator.
实验流程
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).