Red Blood Cell (RBC) Lysis Buffer has been designed, formulated, and tested to ensure
optimal lysis of RBCs in single cell suspensions with minimal effects on leukocytes.
RBCs have to be removed from the whole blood for the study of leukocyte DNA,
proteins or RNA. RBC Lysis Buffer is supplied as a 10X solution and should be diluted
in deionized water prior to use.

1X RBC stock solution contains:
15.5 mM NH4Cl
1.2 mM NaHCO3
0.01 mM EDTA
Ready to use.

Protocol:
Human Whole Blood
1. Add 10 mL of 1X RBC Lysis Buffer per 1 mL of human blood.
2. Incubate for 10-15 minutes at room temperature (no more than 15 minutes).
Note: Observe turbidity to evaluate red blood cell lysis. Once the sample becomes clear, lysis is complete.
3. Centrifuge at 500 x g for 5 minutes at room temperature. Decant supernatant.
4. Resuspend the pellet in the appropriate volume of Flow Cytometry Staining Buffer or buffer of choice.
5. Perform a cell count and viability analysis.
6. Proceed with cell staining or culture, as desired.

Mouse/Rat Spleen or Bone Marrow Cells
1. Harvest tissue and prepare a single-cell suspension.
2. Pellet the cells by centrifugation at 500 x g for 5 minutes at room temperature and decant the supernatant.
3. Resuspend the pellet in 3–10 mL of 1X RBC Lysis Buffer.
4. Incubate for 4–5 minutes at room temperature.
5. Stop the lysis reaction by adding 20–30 mL of 1X PBS.
6. Centrifuge immediately at 500 x g for 5 minutes at room temperature. Decant the supernatant.
7. Resuspend cells in 2 mL of Flow Cytometry Staining Buffer or buffer of choice and centrifuge as in Step 6. Decant supernatant.
8. Resuspend cells in an appropriate volume of Flow Cytometry Staining Buffer or buffer of choice.
9. Perform a cell count and viability analysis.
10. Proceed with cell staining or cell culture, as desired.