Products

Caspase™ Apoptosis Fluorometric Assays

The CasPASE™ Apoptosis assays are designed to monitor apoptosis by measuring various caspase

(protease) activities, a key early indicator of apoptosis in mammalian cells. The assay provides a simple

and easy to follow method that can be monitored with a fluorescence reader.

The assay is based on the detection of cleavage of a synthetic substrate, which has 7-amino-4-

trifluromethyl coumarin (AFC) at the C-terminal. When liberated from the peptide, AFC produces an optical

change that exhibits a fluorescence spectral shift between the substrate-conjugate and the free dye. The

AFC substrate is fluorogenic (detected at 510 to 550 nm with a fluorometer). During the reaction, the substrate

releases AFC free dye and undergoes a fluorescence shift.

The reaction is selectively and irreversibly inhibited by the peptide Z-VAD-FMK (fluoromethyl ketone).

Comparison of the fluorescence from an induced/apoptotic sample with an uninduced control allows one to

determine the fold-increase in protease activity. The Caspase activity can also be quantitated by using a

standard curve established with an appropriate free dye.

When measuring fluorescence, the assay reaction is excited at 380 to 400nm and emission is read at 510 to

550nm.

The assays can be conveniently adapted for high-throughput 96-well format. The assay system may be

used with purified enzyme preparations, cell extracts or tissue lysates.

CasPASE™ assays are available for caspase enzymes 1 through 10 and 13. Each CasPASE™ assay kit is

supplied with necessary assay buffers, enzyme specific AFC-substrate, free dye (AFC), and the potent

caspase inhibitor Z-VAD-FMK for establishing proper positive and negative controls and standards.

Colorimetric Caspase Assays are also available.

Features

• Assays for caspase enzymes 1 through 10 and 13 are available

• Exhibit fluorescence spectral shift

• Conveniently adaptable for high-throughout 96-well format

• All kits supplied with enzyme specific AFC-substrate, free dye (AFC) & inhibitor Z-VAD-FMK

Applications

• Monitor apotosis by measuring various caspase activities in cells and tissues

• Use with purified enzyme preparations, cell extracts, and tissue lysates

• Characterization of caspase enzymes

References

1. Rao, S. et al ( 2016) Evaluation of the anticancer potential of coffee beans: An in vitro study.indian journal of traditional knowlege 15:266.
2. Kulkarni, A et al (2015) IAJPR. 5(2): 937
3. Qi, J. H. et al (2014) Apoptosis. 20:523
4. Timsina, B. et al (2014) Bangladesh J Pharmacol. 9:628
5. Li, W. et al (2014) PLOS. DOI: 10.1371/journal.pone.0094079
6. Bhattacharya, A. et al (2013) Carcinogenesis. 34:2593
7. Li, Q.Q et al (2013) Anticancer Res. 33:1421
8. Vaibhav, K. et al (2013) Neurol. Sci. 34:1321
9. Li, Q.Q et al (2013) Med. Oncol. 30:424
10. Li, L. et al (2012) Cell Stress Chaperon. 18:333
11. Li, C. et al (2013) J. Therm. Biol. 38:513
12. Shrivastava, P. et al (2013) J. Nutr. Biochem. 24:680
13. Lee, R.X. et al (2012) Anticancer Res. 32:3103
14. Soong, G. et al (2012) J Infect Dis 10:1093
15. Sahu, P.P. et al (2012) Mol. Biotech. 52:140
16. Yadav, V. et al (2012) PLOS. DOI: 10.1371/journal.pone.0047796
17. Robbins, D. et al (2012) FEBS Lett. 23:4108
18. Manjoo, A. et al (2010) J. Ortho. Trauma. 24:526
19. Li, Q.Q. et al (2010) Basic Clin. Pharmacol. 107:868
20. Wang, F. et al (2010) PLOS. DOI: 10.1371/journal.pone.0013459
21. Singh, M. and Singh, N. (2009) Mol. Cell. Biochem. 325:107
22. Su, J. et al (2008) Virology. 375:48
23. Khan, T.H. and Sultana, S. (2006) Toxicology. 217:206
24. Yousuf, S. et al (2007) Brain res. 1147:218
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31. Kumi-Diaka, J. and Butler, A. (2000) Biol. Cell. 92:115

Instruction sheet (pdf)

Catalog No.

Product Name

Size

786-200A

 Caspase™ 1, 4, 5 assay with Ac-WEHD-AFC substrate

50 assays

786-200B

 Caspase™ 1, 4, 5 assay with Ac-WEHD-AFC substrate

100 assays

786-201A

 Caspase™ 2 assay with Ac-VDVAD-AFC substrate

50 assays

786-201B

 Caspase™ 2 assay with Ac-VDVAD-AFC substrate

100 assays

786-202A

 Caspase™ 3, 7, 10 assay with Ac-DEVD-AFC substrate

50 assays

786-202B

 Caspase™ 3, 7, 10 assay with Ac-DEVD-AFC substrate

100 assays

786-203A

 Caspase™ 6 assay with Ac-VEID-AFC substrate

50 assays

786-203B

 Caspase™ 6 assay with Ac-VEID-AFC substrate

100 assays

786-204A

 Caspase™ 8 assay with Ac-LETD-AFC substrate

50 assays

786-204B

 Caspase™ 8 assay with Ac-LETD-AFC substrate

100 assays

786-205A

 Caspase™ 9 assay with Ac-LEHD-AFC substrate

50 assays

786-205B

 Caspase™ 9 assay with Ac-LEHD-AFC substrate

100 assays

786-206A

 Caspase™ 13 assay with Ac-LEED-AFC substrate

50 assays

786-206B

 Caspase™ 13 assay with Ac-LEED-AFC substrate

100 assays