Materials

1. Human carcinoma cells (MCF-7, from American Type Culture Collection, Manassas, VA).

2. Buffy coat (leukocyte fraction collected from peripheral blood available from a Red Cross or a Blood Bank).

3. Trypan blue (Invitrogen).

4. Eagle medium (Invitrogen).

5. Accutase (Innovative Cell Technologies).

6. Hank's balanced salt solution (HBSS; JRH Biosciences).

7. Phosphate-buffered saline (PBS; JRH Biosciences).

8. Ethylene diamine tetraacetic acid (EDTA; Invitrogen Corporation).

9. Bovine serum albumin (BSA; Invitrogen Corporation).

10. Accuprep (Accurate Chemicals and Scientific Corporation).

11. Anti-HEA fluorescein isothiocyanate (Miltenyi Biotec, cat. no. 130080-301).

12. Rabbit-anti FITC phosphatase alkaline (cat. no. A4843; Sigma, St. Louis, MO).

13. Fast Redâ„¢ TR-Naphtol AS-MX (cat. no. F-4846; Sigma, St. Louis, MO).

14. Distilled water.

15. Polycarbonate membranes (Whatman; cat. no. 7060-1313).

16. Membrane holders (Swinnex; Millipore, cat. no. SX0001300).

17. Labeling buffer: PBS supplemented with 0.5% bovine serum albumin and 2 ml ethylene diamine tetraacetic acid.

18. Syringe pumps.

19. Particle counter (Beckmann Coulter).

20. Capillary centrifuge.

21. Capillary tubes.

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