1x RBC Lysis Buffer

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. Nucleated RBCs are not effectively lysed with ammonium chloride. This 1x RBC Lysis Buffer is ready to use.

Components

• NH₄ Cl (155 mMol/L)
• KHCO₃ (10 mMol/L)
• EDTA (0.1 mMol/L)

Protocol

1. Ensure that the RBC Lysis Buffer is cold (4-8°C) .
2. Dilute one volume of cell suspension with 10 volumes of 1× Red Blood Cell Lysis Solution.
3. When working wih blood samples:
   Vortex for 5 seconds and incubate for 10-15 minutes (no more than 15 minutes) at 4-8°C.
   
   When working with tissue samples:
   Vortex for 5 seconds and incubate for 2 minutes at 4-8°C.
   Note: Observe turbidity to evaluate red blood cell lysis. Once the sample becomes clear red, lysis is complete.
4. Vortex for 5 seconds and centrifuge at 300×g for 10 minutes at room temperature.
5. Aspirate supernatant completely.
6. Resuspend the cell pellet in an appropriate buffer and proceed to further applications.