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Erythrocyte Lysis

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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 4C 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 - 8C.
  6. 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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