Induction of Apoptosis (Death Receptor Protocol)

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Induction of Apoptosis (Death Receptor Protocol)

Detection Methods of Apoptosis

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Apoptosis in Fas or TNF receptor positive cells can be induced by the appropriate ligands or by using an agonist antibody. A basic protocol using the anti-Fas monoclonal antibody (EOS9.1 catalog # NBP2-00463) is described below.

METHODS

1. Apply 0.05-0.1 µg/ml anti-Fas antibody to 0.5 × 10⁵ cells/ml in fresh media (e.g. Jurkat in fresh RPMI-1640 medium containing 10% fetal bovine serum). Cells should be in the exponential (log) phase prior to treatment. For a negative control incubate untreated cells under identical conditions.
2. Incubate for 3–16 hours in a 37°C incubator. The optimal time should be determined empirically.
3. Harvest the cells by centrifugation.
4. Wash in 1X PBS and resuspend in 1X PBS.
5. Proceed to apoptosis detection .

Learn About Methods to Detect Apoptosis