RBC Lysis
Erythrocyte Lysing Solution (no preservative) 10X Stock Solution
Dissolve in 1 liter of H2O:
- 89.9 g NH4Cl
- 10.0 g KHCO3
- 370.0 mg tetrasodium EDTA
- Adjust pH to 7.3. Store at 4°C in full, tightly closed 50 ml tubes.
1X Working Solution
Add 5 ml 10X Lysing Stock Solution to 45 ml H2O. Mix well. Store at room temperature for up to one week.
Procedure
- Mix 14 ml Lysing Buffer to 1 ml Blood.
- Incubate for 3 – 5 minutes.
- Centrifuge at 300 x g for 5 minutes at room temperature.
- Aspirate supernatant and resuspend pellet in 5 ml of COLD Phosphate Buffered Saline (PBS).
- Centrifuge at 300 x g for 5 minutes at 2 – 8°C.
- Aspirate supernatant and resuspend pellet in 1 ml of COLD PBS with protein carrier.