Overview

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

    Caspase 1 Assay Kit (Fluorometric) (ab39412) provides a simple and convenient method for detecting the activity of caspase 1, which recognizes the sequence YVAD. The assay is based on detection of cleavage of substrate YVAD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). YVAD-AFC emits blue light (Em=400 nm); upon cleavage of the substrate by caspase-1 or related caspases, free AFC emits a yellow-green fluorescence (Em=505 nm), which can be quantified using a fluorometer or a fluorecence microtiter plate reader. Comparison of the fluorescence from a treated sample with an untreated control allows determination of the fold increase in caspase 1 activity.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Caspase 1 (ICE, IL-1beta Converting Enzyme) is the prototypical member of the ICE family of proteases/caspases. Caspase 1 was first identified as a novel protease that generates the proinflammatory cytokine, interleukin 1 beta (IL-1 Beta), by cleaving the pro-interleukin after the Asp116 residue. In addition to its role in the activation of proinflammatory cytokines, Caspase 1 also appears to have functions in some, but not all, types of apoptosis in mammalian cells.

     

  • Tested applicationsSuitable for: Functional Studiesmore details

Properties

  • RelevanceCaspases are a family of cysteine proteases that are key mediators of programmed cell death or apoptosis. The precursor form of all caspases is composed of a prodomain, and large and small catalytic subunits. The active forms of caspases are generated by several stimuli including ligand-receptor interactions, growth factor deprivation and inhibitors of cellular functions. All known caspases require cleavage adjacent to aspartates to liberate one large and one small subunit, which associate into a2b2 tetramer to form the active enzyme. Caspase 1 is similar to the cell death gene CED3 of C. elegans and regulates multiple proinflammatory cytokines, including Interleukin 1b and interferon-gamma-inducing factor. Caspase 1 plays a role in down stream of Caspase 8 which is involved in Fas-mediated apoptosis.
  • Cellular localizationCytoplasmic
  • Alternative names
    • CASP1
    • CASP1 nirs variant 1
    • Caspase 1 apoptosis related cysteine peptidase
    • caspase 1, apoptosis-related cysteine peptidase (interleukin 1, beta, convertase)
    • ICE
    • ICE-like protease
    • IL1 B convertase
    • IL1 beta converting enzyme
    • IL1BC
    • IL1BCE
    • Interleukin 1 B converting enzyme
    • Interleukin 1 beta convertase
    • P45
    see all

Applications

Our Abpromise guarantee covers the use of ab39412 in the following tested applications.

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

Application Abreviews Notes
Functional Studies Use at an assay dependent concentration.

Caspase 1 Assay Kit (Fluorometric) images

  • Titration of the caspase 1 (ab39901) (background signal subtracted, duplicates; +/- SD).

Protocols

References for Caspase 1 Assay Kit (Fluorometric) (ab39412)

This product has been referenced in:
  • Badding MA  et al. Sintered indium-tin oxide particles induce pro-inflammatory responses in vitro, in part through inflammasome activation. PLoS One 10:e0124368 (2015). Functional Studies . Read more (PubMed: 25874458) »
  • Aruna R  et al. Rutin modulates ASC expression in NLRP3 inflammasome: a study in alcohol and cerulein-induced rat model of pancreatitis. Mol Cell Biochem 396:269-80 (2014). Read more (PubMed: 25060908) »

See all 3 Publications for this product

Product Wall

The answers to your questions are as follows:


For doing the caspase-1 assay the cells are lysed and then treated with the substrate. Thus, cell permabilization is not an issue.
A reducing environment is required for the caspases to re...

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The lysis buffer in the kit does not contain any protease inhibitors. The assay relies on keeping the samples on ice to minimize degradation, until the assay at 37C. You could add protease inhibitors to a portion of you lysates set aside for western bl...

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ab39412 has sensitivity of 0.05 - 0.5 ng.

There is 1% Triton-x-100 in the lysis buffer. For best results we recommend using the cell lysis buffer provided. If absolutely necessary for the virus, you may use the RIPA buffer with this kit, but results may vary.

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Unfortunately we do not have a sensitivity range determined for this kit. Fold-increase in Caspase-1 activity can be determined by comparing the results of induced samples with the level of the untreated control. Please let me know if yo...

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Thank you for your inquiry. This result would depend on a number of factors including the cell type in which the Capase-1 activation is being studied, the levels of Caspase-1 activation after induction of apoptosis, time-course for Caspase-1 act...

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Thank you for your call today. I have heard back from the lab about your questions: 1) For tissues, homogenize 10 mg of tissue in 200 ul of lysis buffer. Spin out debris and add 50 ul of supernatant into a well of the 96-well plate then proceed from...

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Thank you for your enquiry. The colorimetric assay has sensitivity of about 5 ng and fluorometric is about 10-100 fold more sensitive. Fluorometric assay is recommended for tissue samples. I hope this information helps. Please do not hesitate to...

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