You have changed your country from  to  . Please be aware that this will change the currency in the purchasing process.

Overview

  • Product nameCaspase 3 Assay Kit (Colorimetric)See all Caspase 3 kits ...
  • Tests
    100 x 1 assay
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay typeEnzyme activity
  • Assay time
    2h 00m
  • Product overview

    Abcam's Caspase 3 Assay Kit (Colorimetric) provides a simple and convenient means for assaying the activity of caspases that recognize the sequence DEVD. The assay is based on spectrophotometric detection of the chromophore p-nitroaniline (p-NA) after cleavage from the labeled substrate DEVD-pNA. The p-NA light emission can be quantified using a spectrophotometer or a microtiter plate reader at 400- or 405 nm. Comparison of the absorbance of p-NA from an apoptotic sample with an uninduced control allows determination of the fold increase in CPP32 activity.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells.

  • Tested applicationsFunctional Studies more details

Properties

  • RelevanceThe caspase family of cysteine proteases play a key role in apoptosis. Caspase 3 (also known as CPP32, YAMA and apopain) is the most extensively studied apoptotic protein among caspase family members. Caspase 3 is synthesized as an inactive pro enzyme that is processed in cells undergoing apoptosis by self proteolysis and/or cleavage by other upstream proteases (e.g. Caspases 8, 9 and 10). The processed form of Caspase 3 consists of large (17kD) and small (12kD) subunits which associate to form an active enzyme. Caspase 3 is cleaved at Asp28 - Ser29 and Asp175 - Ser176. The active Caspase 3 proteolytically cleaves and activates other caspases (e.g. Caspases 6, 7 and 9), as well as relevant targets in the cells (e.g. PARP and DFF). Alternative splicing of this gene results in two transcript variants which encode the same protein. In immunohistochemical studies Caspase 3 expression has been shown to be widespread but not present in all cell types (e.g. commonly reported in epithelial cells of skin, renal proximal tubules and collecting ducts). Differences in the level of Caspase 3 have been reported in cells of short lived nature (eg germinal centre B cells) and those that are long lived (eg mantle zone B cells). Caspase 3 is the predominant caspase involved in the cleavage of amyloid beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease.
  • Cellular localizationCytoplasmic
    • Alternative names
        ApopainApopain precursor CASP 3
        CASP3Caspase 3 apoptosis related cysteine proteaseCaspase 3 p12 subunitCaspase 3 p17 subunitCaspase3CPP 32CPP32CPP32BCysteine protease CPP32Human cysteine protease CPP32 isoform alpha mRNA complete cdsICE3LICEPARP cleavage proteaseSCA 1SCA1SREBP cleavage activity 1Yama protein
      see all

    Applications

    Our Abpromise guarantee covers the use of ab39401 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Notes
    Functional Studies FuncS

    Caspase 3 Assay Kit (Colorimetric) images

    • Caspase 3 Assay Kit (Colorimetric) (ab39401)
      Caspase 3 Assay Kit (Colorimetric) (ab39401)
      H9c2 cells were either untreated (control) or treated with 2µg/ml of recombinant lipocalin-2 (Lcn2) for 24h. Cell lysates were assayed for caspase-3 activity (n = 3, *, p<0.05).

      Image obtained from Xu G et al., JBC, 2012 Feb 11;28(7):4808-17

    Protocols

    References for Caspase 3 Assay Kit (Colorimetric) (ab39401)

    This product has been referenced in:
    • Xu G  et al. Lipocalin-2 induces cardiomyocyte apoptosis by increasing intracellular iron accumulation. J Biol Chem 287:4808-17 (2012). Read more (PubMed: 22117066) »
    • Sandgren P & Stjepanovic A High-yield platelet units revealed immediate pH decline and delayed mitochondrial dysfunction during storage in 100% plasma as compared with storage in SSP+. Vox Sang 103:55-63 (2012). Read more (PubMed: 22220692) »

    See all 2 Publications for this product

    Publishing research using ab39401 ? Please let us know so that we can cite the reference in this datasheet

    Product Wall

    Submit an Abreview for ab39401 Submit a Question for ab39401

    Displaying 1 - 10 of 11 results for Abreviews and Q&A

    10
    Votes: Highest First

    The datasheet for this product says that it can be used for tissue extracts and cell lysates. We have fresh frozen ground squirrel (Ictidomys tridecemlineatus) brain tissue (flash frozen in liquid nitrogen), currently stored at -80 degrees.

    Is ...

    Read More

    Freezing and thawing may partially degrade caspase activity but the protocol, in step 1.2., suggests it will not be a problem: "Transfer supernatant (cytosolic extract) to a fresh tube and put on ice for immediate assay or aliquot and store at -80°C for f...

    Read More

    Inquiry: I'd like to know if the Caspase 3 Assay Kit (Colorimetric) (ab39401) is suitable for bovine cells. Price? Thanks a lot in advance!


    ab39401 should recognize all mammalian forms of Caspase 3. One kit is $470, which provides enough solution for one 96 well plate.

    I want to assay caspase 3 activity in platelet rich plasma (PRP) during storage. Do i use abcam caspase 3 colorimetric assay kit for this purpose? Lysis buffer in the kit is suitable for platelet lysis ? If you have any protocol to assay platelet caspase ...

    Read More

    This kit is good just for cell and tissue lysates but not plasma. If the you are able to collect the platelets from the plasma in a clean manner you will be able to use the kit with the recommended protocol.

    The customer asking if this kit can work on serum or not?
    Caspase 3 Assay Kit (Colorimetric) (ab39401)
    I checked all caspase 3, and all of them in Abcam bot written to work on Serum?
    Truly yours,

    Thank you for contacting us.

    This kit has not been tested with serum sample yet, so we can provide Abtrial discount if customer is interested in testing this kit.

    Please note that this product is "for research use only and are not i...

    Read More

    Dear Technical Team,

    Hope this e-mail finds you well!

    Since, it is not mentioned in the protocol of Caspase 3 Assay Kit (Colorimetric) (ab39401). Kindly suggest us on how to proceed with tissues or how to make single cell suspension as ...

    Read More

    Thank you for contacting us and your interest in our products.

    The Caspase 3 Assay Kit (Colorimetric) (ab39401) can be used with tissue samples and we would suggest following the following protocol:

    1. Induce apoptosis in tissue (10-100)...

    Read More

    Dear.

    I am writing to you because I am interested in the ab39401.
    We investigated the caspase-3 activity inin vivo.
    So, I wonder if the celllysisbuffer of ab39401couldapply tomouse tissue for our study.

    If you have any ques...

    Read More

    Thank you for your enquiry.

    I am pleased to confirm that this kit (ab39401 Caspase 3 Assay Kit (Colorimetric))is compatible with mouse samples. Here are the references we have from this kit. I hope this will provide some reassurance that this k...

    Read More

    Hello, Abcam Tech Support,   I would like to know bout the following issues for my purchase of this kit, Caspase 3 assay kit, ab39401.   1 Does this kit recognize rat caspase-3? 2 Does this kit recognize both activated and non-activated cas...

    Read More

    Thank you for contacting us. I have answered each of your questions below. Please let me know if you have any additional questions about this or any Abcam product.

    1. Does this kit recognize rat caspase-3?

    This will recognize all mam...

    Read More

    What step is the Dilution buffer used for? Is it for diluting the samples for the protein quantification step?

    Thank you for contacting Abcam.

    The dilution buffer is to dilute the final samples before reading their absorbance, in case of the undiluted readings being above the detection range of the instrument.
    Hope this information has been helpful fo...

    Read More

    I order your ab39401 caspase 3 assay kit.
    In the kit the is a dilution buffer, but in the protocol the dilution
    buffer is not mentioned.
    I suppose it is to dilute lysate protein in step 7 (page 6 in
    instruction for use)? Or there is an...

    Read More


    The dilution buffer is used in case you do not have a plate reader an need to read the concentration manually in a cuvette: Step10. "Read samples at 400 or 405 nm in a microtiter plate reader, or
    spectrophotometer using a 100 μl micro-quartz...

    Read More

    je vous remercie,j'attendrai votre réponse.

    Je viens de recevoir une réponse assez vague du laboratoire :
    1.000 ou 10.000 cellulessont peut-être suffisantes si le niveau d'expression de la Caspase 3 par ces cellules est élevé. Le moyen de le savoir est d'essayer.

    

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"