Address: 45-1 Ramsey Road, Shirley, NY 11967, USA    USA: 1-631-559-9269  1-631-448-7888   Fax: 1-631-938-8127  Europe: 44-207-048-3343   Email: info@creative-biomart.com
Home > Products > Assay Kits > Cellular Kits > Caspase-8 assay Kit for drug discovery

Caspase-8 assay Kit for drug discovery

   Share
"Caspase" Related Products.
Cat.No.:
Kit-0172
Product Overview:
The CASPASE-8 Drug Discovery Kit is a complete assay system designed to screen caspase-8 inhibitors. It contains both a colorimetric substrate (IETD-pNA) and a fluorogenic substrate (IETD-AMC). Cleavage of the p-nitroanilide (pNA) from the colorimetric substrate increases absorption at 405nm. The fluorescent assay is based on the cleavage of 7-amino-4-methylcoumarin (AMC) dye from the C-terminus of the peptide substrate. Cleavage of the dye from the substrate increases its fluorescence intensity at 460 nm. The assays are performed in a convenient, 96-well microtiter-plate format. The kit is useful to screen inhibitors of caspase-8, a potential therapeutic target. An inhibitor, IETD-CHO (aldehyde), is included for use as a control.
Size:
96 wells
Description:
Caspase-8 (also known as FLICE, MACH and Mch5) is a member of the interleukin-1β converting enzyme (ICE) family of cysteine proteases. It is implicated as the apical signaling protease in Fas-induced apoptosis. The enzyme is composed of 18 and 11 kDa subunits derived from a common proenzyme. The N-terminal prodomain of the zymogen comprises two "death effector domains" homologous to those of FADD (Mort1). Activation of the Fas-receptor (Apo-1/CD95), by ligand binding, causes FADD to bind the receptor. Procaspase-8 binds the receptor-bound FADD, presumably via interactions between their homologous domains. Binding of the proenzyme leads to its proteolytic processing and activation, possibly by autocatalysis or by interaction with a related caspase (e.g. FLICE2/Mch4). An event which lies downstream from caspase-8 activation is proteolytic activation of the caspase-3 proenzyme, possibly through direct cleavage by caspase-8. The substrate IETD-pNA is based on the recognition sequence for proteolytic activation in procaspase-3. Those tetrapeptide substrates, which incorporate the IETD sequence, are among those cleaved most efficiently by caspase-8.
Storage:
Store all components except the microplate at -70°C for the highest stability. The caspase-8 enzyme component must be handled particularly carefully in order to retain maximal enzymatic activity. Defrost it quickly in a RT water bath or by rubbing between fingers, then immediately store on an ice bath. The remaining unused enzyme should be quickly refrozen by placing at -70°C. The enzyme is stable to freeze/thaw cycles x 4. To minimize the number of freeze/thaw cycles, aliquot the caspase-8 into separate tubes and store at -70°C.
Kit Components:
1. Caspase-8 enzyme (human) (recombinant)FORM: 100 U/µl in assay buffer. One U=1 pmol/min at 30°C, 200 µM IETD-pNA. Purity >95% by SDS-PAGESTORAGE: -70°C; AVOID FREEZE/THAW CYCLES!QUANTITY: 5000 U (50 µl) 2. pNA SUBSTRATE (Ac-IETD-pNA; MW=639)FORM: 2 mM (1.3 mg/ml) in assay bufferSTORAGE: -70°CQUANTITY: 1 ml 3. CALIBRATION STANDARD (p-nitroaniline;MW=138)FORM: 50 µM in assay buffer. A405nm=0.525 cm-1STORAGE: -70°CQUANTITY: 1 ml 4. AMC SUBSTRATE (Ac-IETD-AMC; MW=674)FORM: 0.75 mM (0.5 mg/ml) in assay buffer STORAGE: -70°CQUANTITY: 1 ml 5. AMC CALIBRATION STANDARD (7-amino-4-methylcoumarin; MW=175)FORM: 30 µM in assay bufferSTORAGE: -70°CQUANTITY: 1 ml 6. INHIBITOR (Ac-IETD-CHO; MW=502.5)FORM: 0.1 mM (0.05 mg/ml) in DMSO (dimethylsulfoxide)STORAGE: -70°CQUANTITY: 50 µl 7. ASSAY BUFFER (50 mM HEPES, pH 7.4, 100 mM NaCl, 0.1% CHAPS, 10 mM DTT, 1 mM EDTA, 10% glycerol)STORAGE: -70°CQUANTITY: 20 ml 8. 1/2 -VOLUME MICROPLATE1 clear, 96-wellSTORAGE: Room temperature


Online Inquiry    
Note: There will be extra charge for optional service!
Optional requirements on this protein +Expand
C-fusion   N-fusion  Non-tagged
His   GST  Fc  Others
<1.0 eu/μg   <0.1 eu/μg  <0.01 eu/μg  Not required
Monomer Isolation   Dimer Isolation   Not required
>80% by SDS-PAGE   >90% by SDS-PAGE  >95% by SDS-PAGE  Others
Please input "biomart" as verification code.
Please review Creative Biomart's privacy policy for more information.
Price does not include shipping and packaging costs.Request quote for final price.

If you have any other questions, please call
+1-631-559-9269.

online inquiry

Welcome! For price inquiries, please feel free to contact us through the form below. We will get back to you as soon as possible.

45-1 Ramsey Road, Shirley, NY 11967, USA
USA: 1-631-559-9269  1-631-448-7888
Europe: 44-207-048-3343
FAX: 1-631-938-8127
EMAIL: info@creative-biomart.com

facebook twitter linkedin google plus google plus

Join 10 other subscribers

24x7
CUSTOMER SERVICE
Easy access to products and services you need from our library via powerful searching tools
Copyright © 2007 - 2016 Creative Biomart. All Rights Reserved.