Erythrocyte 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

1. Mix 14 ml Lysing Buffer to 1 ml Blood.
2. Incubate for 3 - 5 minutes.
3. Centrifuge at 300 x g for 5 minutes at room temperature.
4. Aspirate supernatant and resuspend pellet in 5 ml of COLD Phosphate Buffered Saline (PBS).
5. Centrifuge at 300 x g for 5 minutes at 2 - 8°C.
6. Aspirate supernatant and resuspend pellet in 1 ml of COLD PBS with protein carrier.