Caspase 3 Assay试剂盒(Fluorometric) (ab39383)

概述

  • 产品名称
    Caspase 3 Assay试剂盒(Fluorometric)
    参阅全部 Caspase-3 试剂盒
  • 样品类型
    Tissue Extracts, Cell Lysate
  • 检测类型
    Enzyme activity
  • 检测时间
    2h 00m
  • 产品概述

    Caspase 3 Assay Kit (Fluorometric) (ab39383) provides a simple and convenient means for assaying the DEVD-dependent caspase activity. The assay is based on detection of cleavage of substrate DEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). DEVD-AFC emits blue light (λ max = 400 nm); upon cleavage of the substrate by CPP32 or related caspases, free AFC emits a yellow-green fluorescence (Ex/Em = 400/505 nm), which can be quantified using a fluorometer or a fluorescence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-3/CPP32 activity.


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


    Visit our FAQs page for tips and troubleshooting.

  • 说明

    Activation of ICE-family proteases/caspases initiates apoptosis or other cellular processes in mammalian cells.

  • 经测试应用
    适用于: Functional Studiesmore details

性能

  • 功能
    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.
  • 组织特异性
    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.
  • 序列相似性
    Belongs to the peptidase C14A family.
  • 翻译后修饰
    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.
  • 细胞定位
    Cytoplasm.
  • Information by UniProt
  • 别名
    • A830040C14Rik
    • Apopain
    • 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
    • CC3
    • CPP-32
    • CPP32
    • CPP32B
    • Cysteine protease CPP32
    • EC 3.4.22.56
    • LICE
    • mldy
    • OTTHUMP00000165052
    • OTTHUMP00000165053
    • OTTHUMP00000165054
    • PARP cleavage protease
    • Procaspase3
    • Protein Yama
    • SCA 1
    • SCA-1
    • SREBP cleavage activity 1
    • Yama
    see all

应用

Our Abpromise guarantee covers the use of ab39383 in the following tested applications.

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

应用 Ab评论 说明
Functional Studies Use at an assay dependent concentration.

Caspase 3 Assay Kit (Fluorometric) 图像

  • Caspase-3 activity in Jurkat lysates (6.6 x10e5 cells) following 20 hour exposure to 2 uM Camptothecin (ab120115) or 10 ng/mL anti-Fas Ab (MBL). Background signal subtracted, duplicates +/- SD.

实验方案

Caspase 3 Assay Kit (Fluorometric) (ab39383)参考文献

This product has been referenced in:
  • Zou ZZ  et al. Synergistic induction of apoptosis by salinomycin and gefitinib through lysosomal and mitochondrial dependent pathway overcomes gefitinib resistance in colorectal cancer. Oncotarget 8:22414-22432 (2017). Functional Studies ; Human . Read more (PubMed: 26461472) »
  • Tao CC  et al. Epigenetic regulation of HDAC1 SUMOylation as an endogenous neuroprotection against Aß toxicity in a mouse model of Alzheimer's disease. Cell Death Differ 24:597-614 (2017). Functional Studies . Read more (PubMed: 28186506) »

See all 7 Publications for this product

Product Wall



I am afraid that your protease inhibitors might interefere with the assay. Since Caspase is a cysteine protease it is probably inhibited by most protease inhibitor cocktails. The assay is based on the catalytic clevage of the substrate DEVD-A...

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Thank you for your inquiry. This kit will detect active Caspase-3 from samples. The procasepase-3 (inactive) is proteolytically processed into the active Caspase-3. I hope this information helps.  Please contact us with any other questions.

Unfortunately, this is proprietary information.

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 ca...

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

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