CasPASE™ Apoptosis Fluorometric Assays

CasPASE™ Apoptosis Fluorometric Assays

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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-throughput 96-well format
  • All kits supplied with enzyme specific AFC-substrate, free dye (AFC) & inhibitor Z-VAD-FMK

 


Applications

  • Monitor apoptosis by measuring various caspase activities in cells and tissues
  • Use with purified enzyme preparations, cell extracts, and tissue lysates
  • Characterization of caspase enzymes

 

 

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Protocol
786-200A
786-200B
786-201A
786-201B
786-202A
786-202B
786-203A
786-203B
786-204A
786-204B
786-205A
786-205B
786-206A
786-206B
Material Safety Data Sheet
786-200A
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Technical Literature
Bioassays Handbook
Protein Assay Handbook & Selection Guide  An introduction to protein assays.
Catalog Description Size Price(USD) Qty
786-200A
CasPASE™ 1, 4, 5 assay with Ac-WEHD-AFC substrate 50 Assays
$264.00
786-200B
CasPASE™ 1, 4, 5 assay with Ac-WEHD-AFC substrate 100 Assays
$408.00
786-201A
CasPASE™ 2 assay with Ac-VDVAD-AFC substrate 50 Assays
$264.00
786-201B
CasPASE™ 2 assay with Ac-VDVAD-AFC substrate 100 Assays
$408.00
786-202A
CasPASE™ 3, 7, 10 assay with Ac-DEVD-AFC substrate 50 Assays
$264.00
786-202B
CasPASE™ 3, 7, 10 assay with Ac-DEVD-AFC substrate 100 Assays
$408.00
786-203A
CasPASE™ 6 assay with Ac-VEID-AFC substrate 50 Assays
$264.00
786-203B
CasPASE™ 6 assay with Ac-VEID-AFC substrate 100 Assays
$408.00
786-204A
CasPASE™ 8 assay with Ac-LETD-AFC substrate 50 Assays
$264.00
786-204B
CasPASE™ 8 assay with Ac-LETD-AFC substrate 100 Assays
$408.00
786-205A
CasPASE™ 9 assay with Ac-LEHD-AFC substrate 50 Assays
$264.00
786-205B
CasPASE™ 9 assay with Ac-LEHD-AFC substrate 100 Assays
$408.00
786-206A
CasPASE™ 13 assay with Ac-LEED-AFC substrate 50 Assays
$264.00
786-206B
CasPASE™ 13 assay with Ac-LEED-AFC substrate 100 Assays
$408.00