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    caspase-3-assay-kit-colorimetric-ab39401.pdf

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Cell Biology Apoptosis Intracellular Caspases etc Caspases
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Caspase-3 Assay Kit (Colorimetric) (ab39401)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1)Q&A (11)References (178)

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Caspase-3 Assay Kit (Colorimetric) (ab39401)
  • Caspase-3 Assay Kit (Colorimetric) (ab39401)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 2 hr
  • Sample type: Cell Lysate, Tissue Lysate

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Overview

  • Product name

    Caspase-3 Assay Kit (Colorimetric)
    See all Caspase-3 kits
  • Detection method

    Colorimetric
  • Sample type

    Cell Lysate, Tissue 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 Caspase-3 assay protocol is based on the formation of the chromophore p-nitroaniline (p-NA) by cleavage from the labeled substrate DEVD-pNA. The p-NA can be quantified using a spectrophotometer or a microtiter plate reader reading absorbance 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.


    Caspase-3 assay protocol summary:
    - add samples to wells
    - add reaction buffer and DEVD-p-NA substrate and incubate for 60-120 min at 37ºC
    - analyze with microplate reader

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K106 Caspase-3 Colorimetric Assay Kit. K106-100 is the same size as the 100 test size of ab39401.

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

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

    Other caspase and apoptosis assays

    Review the full set of caspase assays, or the apoptosis assay and apoptosis marker guide.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate reader

Properties

  • 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

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Caspases
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Caspases
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Other proteases
    • Kits/ Lysates/ Other
    • Kits
    • Apoptosis Kits
    • Caspase Assays Kits
    • Caspase 3 assay kits
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Caspases
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
    • Neuroscience
    • Processes
  • 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
    modifications

    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

    Cytoplasm.
  • Target information above from: UniProt accession P42574 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • A830040C14Rik
    • Apopain
    • CASP 3
    • CASP-3
    • CASP3
    • CASP3_HUMAN
    • 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
    • Caspase3
    • CC3
    • CPP 32
    • CPP-32
    • CPP32
    • CPP32B
    • Cysteine protease CPP32
    • EC 3.4.22.56
    • ICE3
    • LICE
    • mldy
    • OTTHUMP00000165052
    • OTTHUMP00000165053
    • OTTHUMP00000165054
    • PARP cleavage protease
    • Procaspase3
    • Protein Yama
    • SCA 1
    • SCA-1
    • SCA1
    • SREBP cleavage activity 1
    • Yama
    • Yama protein
    see all

Associated products

  • Corresponding Antibody

    • Anti-Caspase-3 antibody (ab13847)
    • Anti-Cleaved Caspase-3 antibody (ab2302)
    • Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
    • Anti-Caspase-3 antibody (ab4051)
  • Related Products

    • Z-D(OMe)E(OMe)VD(OMe)-FMK, Cell permeable caspase-3 inhibitor (ab120488)
    • Z-VDVAD-FMK, caspase-2 inhibitor (ab142035)
    • Q-DEVD-OPh, caspase-3 inhibitor (ab142037)
    • PAC-1, caspase-3 activator (ab142074)

Images

  • Caspase-3 Assay Kit (Colorimetric) (ab39401)
    Caspase-3 Assay Kit (Colorimetric) (ab39401)Image from Xu G. et al. J Biol Chem. 2012;287(7):4808–4817. doi:10.1074/jbc.M111.275719. Fig. 2G.
    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 Assay Kit (Colorimetric) (ab39401)
    Caspase-3 Assay Kit (Colorimetric) (ab39401)

    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).

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (178)

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

ab39401 has been referenced in 178 publications.

  • Zhu J  et al. MicroRNA-140-5p regulates the proliferation, apoptosis and inflammation of RA FLSs by repressing STAT3. Exp Ther Med 21:171 (2021). PubMed: 33456538
  • Kwon JS  et al. Characterization of ßARKct engineered cellular extracellular vesicles and model specific cardioprotection. Am J Physiol Heart Circ Physiol 320:H1276-H1289 (2021). PubMed: 33513081
  • Wang Y  et al. Paeoniflorin Sensitizes Breast Cancer Cells to Tamoxifen by Downregulating microRNA-15b via the FOXO1/CCND1/ß-Catenin Axis. Drug Des Devel Ther 15:245-257 (2021). PubMed: 33519190
  • Wu J  et al. Hydrogen sulfide inhibits endoplasmic reticulum stress through the GRP78/mTOR pathway in rat chondrocytes subjected to oxidative stress. Int J Mol Med 47:N/A (2021). PubMed: 33537813
  • Chen H  et al. miR-27a-3p regulates the inhibitory influence of endothelin 3 on the tumorigenesis of papillary thyroid cancer cells. Mol Med Rep 23:N/A (2021). PubMed: 33537832
View all Publications for this product

Customer reviews and Q&As

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11-12 of 12 Abreviews or Q&A

Question

je vous remercie,j'attendrai votre réponse.

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Abcam community

Verified customer

Asked on Feb 24 2012

Answer

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.

Read More

Abcam Scientific Support

Answered on Feb 24 2012

Question

i need senstivity range and Species Reactivity for ab 39401 and AB 3983. i need these kit for rats.

Read More

Abcam community

Verified customer

Asked on Jan 16 2009

Answer

Thank you for your enquiry and your interest in our products. The sensitivity range for the fluorometric substrate (K105) is in the tenths of a µM, while the sensitivity of the colorimetric substrates (K106) is in the single digit µM. These kits can detect Caspase 3 in almost all mammalian cells. I hope this information will be useful for you.

Read More

Abcam Scientific Support

Answered on Jan 16 2009

11-12 of 12 Abreviews or Q&A

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