CHX-ACT Apoptosis Made Easy Kit 

Solutions of Cycloheximide and Actinomycin D to be used as enhancers of death receptor induced apoptosis and a solution of Staurosporine, a wide spectrum apoptosis inducer, as a positive control are included. Apoptosis can be determined by measuring caspase 3 activity, using the Sigma CASP-3F kit in a high throughput format.

Apoptotic processes involve well-orchestrated interactions of cell death receptor (CD95-Fas, TNF-R), death receptor adaptors (FADD, TRADD), caspases, Bcl-2 family members and other proteins. Some cells, such as HeLa, HepG2, NT68, rat mesangial cells, and thyroid follicular cells, are resistant to death receptor mediated apoptosis. Maintaining the state of the resistance often requires de novo protein or RNA synthesis. Therefore, treatment with protein synthesis inhibitors, such as cycloheximide (CHX), or RNA synthesis inhibitor, such as actinomycin D (ActD) can sensitize resistant cells. CHX and ActD can also be used as a tool to investigate the apoptosis inhibitors in these cells.

wet ice

−20°C

Wajant, H., et al., Inhibition of death receptor-mediated gene induction by a cycloheximide-sensitive factor occurs at the level of or upstream of Fas-associated death domain protein (FADD). J. Biol. Chem. 275, 24357, (2000) abstract

Myc, A., et al., Characterization of FAP-1 expression and function in thyroid follicular cells. Endocrinology 140, 5431, (1999) abstract

Guo, Y-L., et al., Correlation between sustained c-Jun N-terminal protein kinase activation and apoptosis induced by tumor necrosis factor-alpha in rat mesangial cells. J. Biol. Chem. 273, 4027, (1998) abstract

Fulda, S., et al., Metabolic inhibitors sensitize for CD95 (APO-1/Fas)-induced apoptosis by down-regulating Fas-associated death domain-like interleukin 1-converting enzyme inhibitory protein expression. Cancer Res. 60, 3947, (2000) abstract

UN 3316 6.1/PG 3

1 mg/mL Staurosporine Solution 50 μL

10 mg/mL Cycloheximide Solution 100 μL

10 mg/mL Actinomycin D Solution 100 μL