• Product name
    Caspase 3 Assay Kit (Colorimetric)
    See all Caspase-3 kits
  • Detection method
  • Sample type
    Cell Lysate
  • Assay type
    Enzyme activity
  • Assay time
    2h 00m
  • Product overview

    Caspase 3 Assay Kit (Colorimetric) (ab39401) 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 caspase 3 activity.

    Due to the nature of the substrate, this assay also detects caspase 7 activity.

    Visit our FAQs page for tips and troubleshooting.

  • Notes

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



  • Platform


  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    2X Reaction Buffer 4 x 2ml
    Cell Lysis Buffer 1 x 100ml
    DEVD-pNA 1 x 500µl
    Dilution Buffer 1 x 100ml
    DTT 1 x 400µl
  • Research areas
  • Function
    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.
  • 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.
  • Sequence similarities
    Belongs to the peptidase C14A family.
  • Post-translational
    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.
    S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.
  • Cellular localization
  • Information by UniProt
  • Alternative names
    • A830040C14Rik
    • Apopain
    • CASP-3
    • CASP3
    • Casp3a
    • Caspase 3
    • Caspase 3, apoptosis-related cysteine peptidase
    • Caspase 3, apoptosis-related cysteine protease
    • Caspase 3, apoptosis-related cysteine protease a
    • Caspase-3 subunit p12
    • CC3
    • CPP-32
    • CPP32
    • CPP32B
    • Cysteine protease CPP32
    • EC
    • LICE
    • mldy
    • OTTHUMP00000165052
    • OTTHUMP00000165053
    • OTTHUMP00000165054
    • PARP cleavage protease
    • Procaspase3
    • Protein Yama
    • SCA 1
    • SCA-1
    • SREBP cleavage activity 1
    • Yama
    see all


  • 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

  • Caspase 3 in Jurkat lysates (3.3 x10e6 cells) following 20 hour exposure to 2 uM Camptothecin (ab120115) or 10 ng/mL anti-Fas Ab (MBL).



This product has been referenced in:
  • Lam TG  et al. New metformin derivative HL156A prevents oral cancer progression by inhibiting the insulin-like growth factor/AKT/mammalian target of rapamycin pathways. Cancer Sci 109:699-709 (2018). Read more (PubMed: 29285837) »
  • Leger T  et al. Dietary canolol protects the heart against the deleterious effects induced by the association of rapeseed oil, vitamin E and coenzyme Q10 in the context of a high-fat diet. Nutr Metab (Lond) 15:15 (2018). Functional Studies . Read more (PubMed: 29456586) »

See all 47 Publications for this product

Customer reviews and Q&As

It doesn't work for tissue samples

Inconclusive Good 4/5 (Ease of Use)
We tried to quantify Caspase 3 activity in mouse mammary gland during mammary involution after lactation. We tested different protein amounts fron 50 to 200 microgrames (as indicated in protocol booklet), but the product did not work in any case. We wanted to compare Caspase 3 activity in three samples at different timepoints of involution, and we were not able to observe differences (A), which really exist, as we confirmed by western blot of the same samples (B). We checked the kit was not faulty testig in HC11 mammary cell line, inducing apoptosis with cisplatin 15 ug/ml for 24h (C).
The technical service told us it was a problem with our samples and the did not offer a way to solve our problem.
Then, we found a fluorimetric assay ab39383, which is exactly the same but with a fluorometric substrate instead of a colorimetric one, which is more sensitive as abcam scientific support says in the questions and answer section of the product wall in their website. We aquired the fluorometric substrate and repeated the assay with the same samples. This time, the product worked perfectly (D).
So, for mouse mammary gland tissue samples, I would not recommend this product. It seems it has not enough sensitivity. I would try the fluorimetric assay.

Mr. Adrian Blanco

Verified customer

Submitted Jul 17 2014

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 fo...

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

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.

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 no...

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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 (10-1...

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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 thi...

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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 ...

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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...

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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-qua...

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1-10 of 12 Abreviews or Q&A


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