To provide a phenotypic assay for human monocytes/macrophages.
Author: J.E. Colligan et al, Translator: Xuitao Cao et al. This experiment is from the "Comprehensive Immunology Laboratory Guide".
Operation method
Phenotyping of Human Monocytes/Macrophages Move basic program Materials Single nucleated cells (heparin-anticoagulated blood or heparin-anticoagulated bone marrow) or peripheral blood single nucleated cells (cell 8.1) or mononuclear/macrophage cells (cell 8.5, negative sorting recommended) to be tested PBS PBS/Heparin: PBS with 0.1% (V/V) Heparin Blocking IgG at 3 mg/ml (e.g., normal mouse IgG, Caltag) Fluorescein-labeled monoclonal antibody (Table 6. 3. 2) E M A (Ethidium monoazide) solution ACK lysate (freshly prepared, should be stored at room temperature for < 12 h) 7 2 % (m /V) Formaldehyde 12m m X 75m m test tube 15 ml cone-bottomed plastic centrifuge tubes (Falcon) Low power incandescent lamps (e.g. 40W incandescent) Sorvall R T 6000B Centrifuge and Rotor When using heparinized anticoagulated blood or anticoagulated bone marrow: add approximately IOml of anticoagulated blood or 1-3ml of anticoagulated bone marrow to a 15ml cone-bottomed plastic centrifuge tube and centrifuge at 4°C for 3 minutes at 1500 g. Discard the blood plasma, add 100ml of PBS/heparin, and tightly cap the tube. For more product details, please visit Aladdin Scientific website.
