Materials

1. MiniMACS magnetic isolation system using MS+ column (Miltenyi Biotec, Auburn, CA)

2. NK cell-targeting, primary antibody: mouse anti human CD56-phy-coerythrin (PE) antibody conjugate, isotype IgG1 (Immunotech, Marseille, France).

3. Secondary, magnetic tagging antibody: mouse anti PE-MACS microbeads antibody conjugate, isotype IgG1 (Miltenyi Biotec, Auburn, CA).

4. Ficoll-Hypaque preparation (Accurate Chemical and Scientific Corporation, Westbury, NY).

5. Degassed solution of phosphate-buffered saline (PBS) without calcium and magnesium, supplemented with 0.1% (w/v) bovine serum albumin and 2 ml ethylene diamine tetraacetic acid.

6. Lysis buffer: 154 ml NH4Cl, 10 mM KHCO3, 0.1 ml ethylene diamine tetraacetic acid.

7. Other laboratory equipment and reagents: visible light microscope, hemacytometer, Trypan blue solution, laboratory balance, refrigerated centrifuge, household bleach 10% solution, automated cell counter (Coulter Z1 Counter, Beckman-Coulter, Miami, FL), flow cytometer (EPICS Elite II, Beckman-Coulter).

0 0

Post a comment