用法 :
RBC lysis buffer is supplied as a 10X solution and should be diluted to 1X in deionized water. 100 ml of 10X concentrate will yield a quantity of 1X solution that is sufficient to lyse 500 samples.
The pH of the 1X solution should fall within the range of pH 7.1-7.4. Adjust the pH if necessary. Warm the 1X solution to room temperature prior to use.
NOTE: The blood sample must be <1 month old and been stored in anticoagulants (i.e. EDTA, heparin, and citrate).
PROTOCOL
1. Add whole blood to RBC Lysis Buffer at a ratio of approximately 1 part blood to 2 parts Lysis Buffer. Invert the tube to mix and incubate for 5 minutes at room temperature on a shaking platform or with periodic inversions. Do not vortex.
2. Centrifuge for 5 minutes then remove supernatant carefully without disturbing the pellet.
Note: If pellet is not white, repeat steps 1 and 2 with additional RBC Lysis Buffer.
3. If a lower red pellet is still visible below the white pellet then repeat steps 1 and 2 with fresh blood with the following modifications: a. Ensure the RBC Lysis Buffer is at room temperature. b. Increase the incubation to 15 minutes c. Use a higher ratio of RBC Lysis Buffer to blood (3:1).