Note: It is recommended to dilute the RBC Lysis Buffer (10X) just prior to use.
1. Dilute the RBC Lysis Buffer (10X) to a 1X working concentration by adding 1 part RBC Lysis Buffer (10X) to 9 parts room temperature deionized water. Warm the solution to room temperature.
2. Add 2.0 ml of 1X RBC Lysis Buffer to the prepared sample of whole blood (50-100 µL per tube), gently vortex the sample.
3. Incubate for 10-15 minutes at room temperature protected from light.
4. Centrifuge cells at 500 x g for 5 minutes at room temperature. Carefully aspirate the supernatant without disturbing the cell pellet.
5. Wash the cells once in excess with Flow Cytometry Antibody Dilution Buffer #13616, repeat the centrifugation in step 4.
6. Resuspend the cells in the appropriate volume for analysis.
Note: Above outlines the necessary steps for lysis of human whole blood samples. If working with mouse blood or splenocytes, decrease the incubation time (step 3) to 5 minutes. Further optimization may be required.
The 10X solution is stable for 6 months when stored at 4°C.
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