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Induction of apoptosis in cells

Procedure for biological and chemical induction of apoptosis in cells.
Last edited Tue 14 Feb 2023

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.

Stage 1 - Apoptosis 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. Our mouse monoclonal anti-Fas antibody [UT-1] (ab11881) can be used for apoptosis indication.

Steps

1

Grow Jurkat cells in RPMI-1640 containing 10% fetal bovine serum (FBS) in a humidified 5% CO2 incubator at 37°C.

2

Harvest exponentially growing cells at a concentration of 105 cells/mL by centrifugation at 300–350 x g for 5 min.

  • Resuspend cells in fresh medium to a final concentration of 5 x 105 cells/mL.
3

Add anti-Fas (anti-CD95) mAb to the appropriate concentration. For ab11881, use a final concentration of 2–20 mg/mL.

  • Incubate for 2–4 hr in a 37°C incubator.

Stage 2 - Harvesting cells

Steps

1

Harvest the cells by centrifugation at 300–350 x g for 5 min.

2

Remove all medium and resuspend cells in PBS.

3

Repeat centrifugation step and resuspend cells.

  • In PBS to 1.5 x 106 cells/mL.
4

Proceed to detect apoptosis using your method of choice.

Stage 3 - Detect apoptosis

Steps

1

Collect cells and detect apoptosis using your method of choice.