APT165 | Caspase 3 Colorimetric Activity Assay Kit, DEVD

APT165
100 assays  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey ApplicationsDetection Methods
      MaACTChromogenic
      Description
      Catalogue NumberAPT165
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionCaspase 3 Colorimetric Activity Assay Kit, DEVD
      OverviewActivation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. Chemicon's Caspase-3 Colorimetric Activity Assay Kits provide a simple and convenient means for assaying the activity of caspases that recognize the sequence DEVD. The assay is based on spectophotometric detection of the chromophore p-nitroaniline (pNA) after cleavage from the labeled substrate DEVD-pNA. The free pNA can be quantified using a spectrophotometer or a microtiter plate reader at 405 nm. Comparison of the absorbance of pNA from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-3 activity.
      Alternate Names
      • CPP32
      Materials Required but Not Delivered· Microcentrifuge and 1.5 mL Microcentrifuge tubes

      · 37°C Waterbath or Incubator

      · Spectrophotometer or Microplate Reader

      · 96 well Microtiter Plate
      References
      Product Information
      Components
      • 5X Cell Lysis Buffer (Part No. 90065): 5 mL
      • 5X Assay Buffer (Part No. 90066): 10 mL
      • Caspase-3 Substrate (Ac-DEVD-pNA) (Part No. 90080): 1 mL of 3 mg/mL solution
      • Caspase-3 Inhibitor (Ac-DEVD-CHO) (Part No. 90067): 50 μL of 100 μM (0.05 mg/mL) in DMSO
      • pNA Standard (Part No. 90085): 250 μL of 10 mM in DMSO
      Detection methodChromogenic
      Applications
      ApplicationCaspase-3 Colorimetric Activity Assay Kits provide a simple & convenient means for assaying the activity of caspases that recognize the sequence DEVD.
      Key Applications
      • Activity Assay
      Biological Information
      Species Reactivity
      • Mammals
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein cleaves and activates caspases 6, 7 and 9, and the protein itself is processed by caspases 8, 9 and 10. It is the predominant caspase involved in the cleavage of amyloid-beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease. Alternative splicing of this gene results in two transcript variants that encode the same protein.
      Gene Symbol
      • CASP3
      • apopain
      • procaspase3
      • CPP-32
      • Apopain
      • Yama
      • CASP-3
      • CPP32B
      • SCA-1
      • CPP32
      • EC 3.4.22.56 [Contains: Caspase-3 subunit p17
      • Caspase-3 subunit p12].
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P42574 # Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp- -Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop- helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin.
      SIZE: 277 amino acids; 31608 Da
      SUBUNIT: Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 17 kDa (p17) and a 12 kDa (p12) subunit.
      SUBCELLULAR LOCATION: Cytoplasm.
      TISSUE SPECIFICITY: Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.
      PTM: Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
      SIMILARITY: SwissProt: P42574 ## Belongs to the peptidase C14 family.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStore kit materials at -20°C up to their expiration date.

      Special Precautions:

      · After thawing reagents, use immediately or aliquot and freeze at -20oC for longer storage. Avoid repeated freeze/thaw cycles.

      · The Caspase-3 substrate and pNA standard are especially light sensitive. Maintain these reagents in amber or covered containers.
      Packaging Information
      Material Size100 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Ras signaling in NGF reduction and TNF-?-related pancreatic ? cell apoptosis in hyperglycemic rats.
      Selda Gezginci-Oktayoglu,Sehnaz Bolkent
      Apoptosis : an international journal on programmed cell death  17  2012

      Show Abstract
      21938476 21938476
      Xenon is not superior to isoflurane on cardiovascular function during experimental acute pulmonary hypertension.
      A B Roehl,P Steendijk,R Rossaint,C Bleilevens,A Goetzenich,M Hein
      Acta anaesthesiologica Scandinavica  56  2012

      Show Abstract
      22260254 22260254
      Involvement of oxygen-regulated protein 150 in AMPK-mediated alleviation of lipid-induced endoplasmic reticulum stress.
      Wang Y, Wu Z, Li D, Wang D, Wang X, Feng X, Xia M
      J Biol Chem  2011

      Show Abstract
      21296878 21296878

      User Guides

      Title
      Caspase-3 Colorimetric Activity Assay Kit