Triggers KG-1a cell aggregation
For whole blood samples, add 5 μL Anti-Human CD3-APC/CD4-FITC/CD8a-PE Cocktail to 100 μL anticoagulant-treated blood sample. Mix and incubate the sample at 4°C in the dark for 30 min. Remove red blood cells with RBC lysis solution following the manufacturer's instruction. Wash the cell with cell staining buffer and discard the supernatant after centrifugation at 300 g for 5 min. Resuspend the cells with 200 μL cell staining buffer and load the sample on flow cytometer for detection. For other samples, 1×106 dissociated single cells are centrifuged at 300 g for 5 min with the supernatant discarded. Resuspend the cells with 100 μL cell staining buffer and add 5 μL Anti-Human CD3-APC/CD4-FITC/CD8a-PE Cocktail. Mix and incubate the sample at 4°C in the dark for 30 min. Add cell staining buffer to each tube, centrifuge at 300 g for 5 min and discard the supernatant. Resuspend the cells with 200 μL cell staining buffer and load the sample on flow cytometer for detection.
For whole blood samples, add 5 μL Anti-Human CD3-APC/CD4-FITC/CD8a-PerCP-Cyanine5.5 Cocktail to 100 μL anticoagulant-treated blood sample. Mix and incubate the sample at 4°C in the dark for 30 min. Remove red blood cells with RBC lysis solution following the manufacturer's instruction. Wash the cell with cell staining buffer and discard the supernatant after centrifugation at 300 g for 5 min. Resuspend the cells with 200 μL cell staining buffer and load the sample on flow cytometer for detection. For other samples, 1×106 dissociated single cells are centrifuged at 300 g for 5 min with the supernatant discarded. Resuspend the cells with 100 μL cell staining buffer and add 5 μL Anti-Human CD3-APC/CD4-FITC/CD8a-PerCP-Cyanine5.5 Cocktail. Mix and incubate the sample at 4°C in the dark for 30 min. Add cell staining buffer to each tube, centrifuge at 300 g for 5 min and discard the supernatant. Resuspend the cells with 200 μL cell staining buffer and load the sample on flow cytometer for detection.
For whole blood samples, add 5 μL Anti-Human CD3-FITC/CD16+CD56-PE Cocktail to 100 μL anticoagulant-treated blood sample. Mix and incubate the sample at 4°C in the dark for 30 min. Remove red blood cells with RBC lysis solution following the manufacturer's instruction. Wash the cell with cell staining buffer and discard the supernatant after centrifugation at 300 g for 5 min. Resuspend the cells with 200 μL cell staining buffer and load the sample on flow cytometer for detection. For other samples, 1×106 dissociated single cells are centrifuged at 300 g for 5 min with the supernatant discarded. Resuspend the cells with 100 μL cell staining buffer and add 5 μL Anti-Human CD3-FITC/CD16+CD56-PE Cocktail. Mix and incubate the sample at 4°C in the dark for 30 min. Add cell staining buffer to each tube, centrifuge at 300 g for 5 min and discard the supernatant. Resuspend the cells with 200 μL cell staining buffer and load the sample on flow cytometer for detection.
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