Additional Information
Product Name: | Caspase-8, 10 Activity Fluorometric Assay Kit |
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Also Known As: | N/A |
Catalog No.: | J7110 |
Size: | 1 Kit (250 assays) |
Molecular Weight: | N/A |
Species: | N/A |
Source: | N/A |
Stock: | N/A |
Concentration: | See tube label |
Quality Assurance: | Inhibitor and substrate > 98% by HPLC and NMR |
Storage: | Store at -80°C; avoid multiple freeze-thaw cycles |
PDF Data Sheet: | PDF datasheet, MSDS |
NCBI RefSeq: | N/A |
Image(s): | J7110 image HCT116 cells were treated with 50 ng/ml 6xHis-TRAIL for 4 hours to activate extrinsic apoptosis. 25 μg cell lysates were incubated with (open circles) or without (solid circles) 20 μM Z-VAD-FMK (a pan caspase inhibitor) for 10 min at 37 0C. 25 μM Ac-IETD-AFC (A) or Ac-DEVD-AFC (B) was added to initiate the reaction. AFC fluorescence was recorded with a plate reader using excitation/emission filter set at 400±40/508±30 nm, respectively. |
Shipping Method: | Dry ice shipping |
References: | N/A |
Details
Caspase-8, 10 Activity Fluorometric Assay Kit can be used for assaying caspase-8, 10, 3, 7 activities in cell/tissue extracts in a 96-well plate format. Caspase-8, 10, 3, 7 cleave the supplied fluorogenic substrates. The fluorescence of released AFC can be monitored using a plate reader or a fluorometer at the excitation/emission wavelength of 400 nm/505 nm, respectively. The 96-well format assay can be completed in 2-4 hours.
Ac-IETD-AFC is a fluorogenic substrate of caspase-8/10.
Ac-DEVD-AFC is a fluorogenic substrate of caspase-3/7.
Z-VAD-FMK is a pan caspase inhibitor, used to inhibit caspases in reactions to obtain background signals.
TRAIL-treated HCT116 cell lysates contain activated caspase-8/10/3/7, used as a positive control.
AFC is used for generating a standard curve for quantitating the amount of AFC produced in reactions.
Components
• 500X Ac-IETD-AFC (12.5 mM in DMSO)...................50 µl
• 1000X Ac-DEVD-AFC (25 mM in DMSO)..................25 µl
• 1000X Z-VAD-FMK (25 mM in DMSO).......................25 µl
• TRAIL-treated HCT116 cell lysates (5 mg/ml)............25 µl
• AFC (0.1 mM in DMSO)..............................................250 µl
• Tris (2 M, pH 7.1 at 37°C) ..........................................1 ml
• DTT (200 mM).............................................................1 ml