Background4A8H9 reacts with high affinity receptor for IgG (FcyRI), a 75 kDa type 1 trasmembrane glycoprotein. CD64 is expressed on monocytes and macrophages but not on lymphocytes or resting granulocytes. CD64 play a role in phagocytosis, and antibody dependent cellular cytotoxicity ( ADCC). It also participates in cytokine and superoxide release.
Product Information
IsotypeMouse IgG1
Clone Number4A8H9
Experimental Methods1.Take 100μl peripheral blood anticoagulated by EDTA and add to the bottom of 5ml tube;
2.Add 10μl labeled antibody to the bottom of flow tube mixing with the whole blood, incubate for 20 minutes at room temperature away from light;
3.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);
4.Sample tube is set to 1000 rpm centrifugation for 5 minutes, discard the supernatant;
5.Add 2 ml PBS wash buffer to resuspend the cells, then1000 rpm centrifugation for 5 minutes, discard the supernatant;
6.Add 0.5 ml PBS 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 ℃ then measured).
[PBS wash buffer: PBS +1% FBS +0.1% NaN3; Cat.: FXP005]
[Cell fixation: 2% formaldehyde solution]
Notices1.This reagent has been pre-diluted for use at the recommended Volume per Test.
We typically use 1 X 106 cells in a 100μl experimental sample (a test);
2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing;
4. If the sample can not be timely analysis, please fixed;
5. For research use only, not for diagnostic or therapeutic use.
Related productsE16HU064 Mouse Anti-Human CD64,Purified mAb FCM IF
E16HP064 Mouse Anti-Human CD64, PE Conjugated mAb FCM IF
Profile of peripheral blood lymphocytes analyzed by flow cytometry
References1. Ledbetter, J et al. (1981) J Exp Med 153:310-323.
2. Haynes, B et al. (1986) Leukocyte Typing II: Human T lymphocytes. New York: Sprimger-Verlag; 3-30.
3. Clevers H et al. (1988). Ann Rev Immunol 6: 629-662.
4. Barclay, N et al. (1993). The Leucocyte Factsbook. Academic Press. San Diego.