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Apoptosis may be induced in experimental systems through a variety of methods. In general, methods to induce apoptosis can be divided into two categories: biological induction and chemical induction.
Activation of Fas or TNF receptors by their respective ligands, or by cross-linking with an agonist antibody, induces apoptosis of Fas- or TNF receptor-bearing cells. Here we describe a general protocol to induce apoptosis using an anti-Fas receptor (anti-CD95) monoclonal antibody (mAb) in Jurkat cells. The mouse monoclonal anti-Fas antibody [UT-1] (ab11881) can be used for apoptosis indication.
Apoptosis inducers act on several apoptosis-related proteins to promote apoptotic cell death. Depending on the agent selected and the concentrations used, apoptotic events can be detected between 8–72 h post-treatment. However, not all reagents will affect a particular cell line in the same way.
These are general guidelines for inducing cellular damage with chemical agents that will lead to apoptosis.
Agent | Concentration |
Doxorubicin hydrochloride (ab120629) | 0.2 µg/mL (25 µg/mL stock prepared in H2O) |
Staurosporine (ab120056) | 1 µM (1 mM stock prepared in DMSO) |
Etoposide (ab120227) | 1–10 µM (1 mM stock prepared in DMSO) |
Camptothecin (ab120115) | 2–10 µM (1 mM stock prepared in DMSO) |
Paclitaxel (ab120143) | 50–100 nM (stock prepared in DMSO) |
Vinblastine (ab145649) | 60 nM (stock prepared in methanol) |
Agent | Concentration |
Z-VAD-FMK (ab120382) | 50 µM (stock prepared in DMSO) |