For whole blood samples, add 5 μL Anti-Human CD3-FITC/CD19-APC/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/CD19-APC/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.
For whole blood samples, add 5 μL Anti-Human CD3-FITC/CD19-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/CD19-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-FITC/CD4-PE/CD45-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-FITC/CD4-PE/CD45-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/CD4-PE/CD8a-PE/Cyanine7/CD45-PerCP 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/CD4-PE/CD8a-PE/Cyanine7/CD45-PerCP 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.
Each lot of this antibody is quality control tested by flow cytometric analysis. The amount of the reagent is suggested to be used 5 µL of antibody per test (million cells in 100 µL staining volume or per 100 µL of whole blood). Please check your vial before the experiment. Since applications vary, the appropriate dilutions must be determined for individual use.
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