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Separation of T-cell and B-cell lymphocytes with Combed, Scrubbed Nylon Wool Fibre Combed nylon wool is an ideal separation medium for lymphocytes. Using nylon wool, B cells can be separated from T cells in a simple and quick way for immunological research. Mouse granulocytes and B cells stick to the nylon fibres, while T cells pass through the nylon wool. The nylon wool is in principle ready for use and can be autoclaved or pre-treated if necessary.
For literature see also:
Current Protocols in Immunology (1992) 3.2.1-3.2.4
Karen S. Hathcock: "T Cell Enrichment by Nonadherence to Nylon
Short protocol:
1) Method
• finely pluck 0.5 to 1g of nylon wool, stuff into a 10 ml syringe and autoclave. Wash and equilibrate the nylon wool with HBSS, RPMI 1640 enriched with 10% TCS or T-cell medium
• suspend 1-2 x 10⁸ mononuclear cells in 2 ml medium and pipette into the syringe
• Incubate the syringe at 37 °C for 45 - 60 min.
• Collect and resuspend non-adherent T cells by washing with HBSS. Let the syringe run out slowly - do not press, refill with medium; repeat the procedure if necessary
2) Solutions
• T-cell medium
• RPMI, 5% FKS, L-Glu, NEAA, Pen / Strep, HEPES, SodPyr
• PBS and PBS + 1% TCS
• Lysis buffer
