Shreffler:RBC Lysis Buffer for Eos
RBC Lysis Buffer recipe (for Eosinophil protocol)
- NH4Cl (Sigma A0171-100G)
- KHCO3 (Sigma 237205-100G)
- 0.5 M EDTA (Fisher EC 200-449-4)
- 500 ml millipore filter flask (Millipore SCGPU05RE)
- Measure out 500 ml DI water in beaker (slightly less is OK).
- Bring solution to 0.8% (4 g) NH4Cl (Ammonium Chloride, above Bert’s bench).
- Bring solution to 0.1 mM EDTA (100 µl of 0.5 M EDTA stock).
- Bring pH to 7.2 using 2M KHCO3 (Potassium Bicarbonate, in 50 ml conical above Bert’s bench, but if more see recipe below).
- Filter in 500 ml flask.
- For 2 M KHCO3, add 5 g KHCO3 to 25 ml DI water in 50 ml conical tube.