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작성일 : 17-03-28
[G-Biosciences] Caspase™ Apoptosis Fluorometric Assays

 

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.

 

 

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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
  25. Zhang, A. et al (2006) Am J Physiol Renal Physiol 291:F1332
  26. Adamus, G. et al (2006) J. AutoImmun. 26:146
  27. Wang, G. et al (2005) CMLS. 62:881
  28. Chiou, S. et al (2005) Gastroenterology. 128:63
  29. Dallalio, G. and Means, R.T. (2003) J. Lab. Clin. Med. 141:395
  30. Adamus, G. et al (2003) J. AutoImmun. 21:121
  31. Kumi-Diaka, J. and Butler, A. (2000) Biol. Cell. 92:115

 

 

Instruction sheet (pdf)

 

 

Ordering informations

    

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

    

   

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