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Erythrocyte Lysing
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.
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Dennis J. Young,
UCSD Cancer Center Flow Cytometry Core Facility
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