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Cell Biology Apoptosis Intracellular Caspases etc Caspases
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Caspase-1 Assay Kit (Fluorometric) (ab39412)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (9)References (20)

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Functional assays: Caspase 1 assay kit (ab39412)
  • Functional assays: Caspase 1 Assay Kit (Fluorometric) (ab39412)

Key features and details

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

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Overview

  • Product name

    Caspase-1 Assay Kit (Fluorometric)
    See all Caspase-1 kits
  • Detection method

    Fluorescent
  • Sample type

    Tissue Extracts, Cell Lysate
  • Assay type

    Enzyme 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 caspase-1 assay protocol is based on the 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 (Ex/Em=400/505 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.


    Caspase-1 assay protocol summary:
    - add samples to wells
    - add reaction buffer and YVAD-AFC substrate and incubate for 1-2 hr
    - analyze with a microplate reader

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K110 Caspase-1 Fluorometric Assay Kit. K110-100 is the same size as the 100 test size of ab39412.

    Caspase-1 (ICE, IL-1beta Converting Enzyme) is the prototypical member of the ICE family of proteases/caspases.

    Other caspase and apoptosis assays

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

  • 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
    DTT 1 x 0.4ml
    YVAD-AFC 1 x 0.5ml
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Caspases
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Caspases
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Kits/ Lysates/ Other
    • Kits
    • Apoptosis Kits
    • Caspase Assays Kits
    • Caspase 1 assay kits
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Caspases
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
  • Relevance

    Caspases 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 localization

    Cytoplasmic
  • Alternative names

    • CASP-1
    • CASP1
    • CASP1 nirs variant 1
    • Caspase 1 apoptosis related cysteine peptidase
    • Caspase 1 apoptosis related cysteine protease
    • caspase 1, apoptosis-related cysteine peptidase (interleukin 1, beta, convertase)
    • Caspase1
    • ICE
    • ICE-like protease
    • IL-1 beta-converting enzyme
    • IL-1BC
    • IL1 B convertase
    • IL1 beta converting enzyme
    • IL1B convertase
    • IL1BC
    • IL1BCE
    • Interleukin 1 B converting enzyme
    • Interleukin 1 beta convertase
    • Interleukin 1 beta convertase precursor
    • Interleukin 1B converting enzyme
    • Interleukin-1 beta-converting enzyme
    • P45
    see all

Associated products

  • Related Products

    • Z-LE(OMe)VD(OMe)-FMK, derivative of Z-LEVD-FMK (ab120489)
    • Zaprinast, cGMP phosphodiesterase inhibitor (ab120940)
    • Z-YVAD-FMK, Irreversible caspase-1 inhibitor (ab141388)
    • Z-WEHD-FMK, caspase-1, caspase-5 and cathepsin B inhibitor (ab142027)

Images

  • Functional assays: Caspase 1 assay kit (ab39412)
    Functional assays: Caspase 1 assay kit (ab39412)Image from Badding M.A et al., PLoS One 10(4). Fig 6b. doi: 10.1371/journal.pone.0124368. Reproduced under the Creative Commons license https://creativecommons.org/publicdomain/zero/1.0/

    Badding M.A et al investigated the cytotoxicity of Indium-tin oxide (ITO). Previously,ITO had shown to be cytotoxic in cultured cells and pro-inflammatory in pulmonary animal models. ITO is used to make transparent conductive coatings for touch screens and liquid crystal display electronics. RAW cells were plated at 5 x 105 cells/well and treated with SITO, LPS, or Min-U-sil (cells were first primed with LPS). Cells were washed, lysed, and 100 μg of lysates were assayed using Caspase 1 assay kit (ab39412). PBS was used as a control. All conditions were run in duplicate wells and three independent experiments were performed for each time point.

  • Functional assays: Caspase 1 Assay Kit (Fluorometric) (ab39412)
    Functional assays: Caspase 1 Assay Kit (Fluorometric) (ab39412)

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

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (20)

Publishing research using ab39412? Please let us know so that we can cite the reference in this datasheet.

ab39412 has been referenced in 20 publications.

  • Yin XL  et al. Tojapride prevents CaSR-mediated NLRP3 inflammasome activation in oesophageal epithelium irritated by acidic bile salts. J Cell Mol Med 24:1208-1219 (2020). PubMed: 31859410
  • Parker H  et al. Haematopoietic stem cell gene therapy with IL-1Ra rescues cognitive loss in mucopolysaccharidosis IIIA. EMBO Mol Med 12:e11185 (2020). PubMed: 32057196
  • Mukherjee S  et al. Deubiquitination of NLRP6 inflammasome by Cyld critically regulates intestinal inflammation. Nat Immunol 21:626-635 (2020). PubMed: 32424362
  • Wu L  et al. RIPK3 Orchestrates Fatty Acid Metabolism in Tumor-Associated Macrophages and Hepatocarcinogenesis. Cancer Immunol Res 8:710-721 (2020). PubMed: 32122992
  • Gu C  et al. miR-590-3p Inhibits Pyroptosis in Diabetic Retinopathy by Targeting NLRP1 and Inactivating the NOX4 Signaling Pathway. Invest Ophthalmol Vis Sci 60:4215-4223 (2019). PubMed: 31618425
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Question

I buy the Caspase 1 Assay Kit (Fluorometric) (ab39412), and I have two question about the reactives including on it:

1- The YVAD-AFC substrate is pearmeable into the cells?
2- The DTT is included for what?

Read More

Abcam community

Verified customer

Asked on Jul 08 2014

Answer

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 remain stable in solution and retain activity. This is why DTT is used.

Read More

Sybille Rex

Abcam Scientific Support

Answered on Jul 08 2014

Question

Hello, I was wondering if the lysis buffer that comes with the Casapse-1 activation assay contains protease inhibitors? We typically add those to our lysis buffer for lysates run in western blots. We plan to lyse cells and use part of the lysate for this assay/kit and the rest will be run in western blots. Thank you.

Read More

Abcam community

Verified customer

Asked on Apr 17 2014

Answer

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 blots immediately after homogenization, but do not add them to the portion you intend to use for the assay, as caspase activity could be inhibited.

For a western blot test, I suggest loading a lane in a gel with a sample that did not receive inhibitors, and compare the result to a lane that was loaded with sample + inhbitors, to see if protease inhibitors are necessary to keep caspase 2 intact.

Read More

Tom Ruyle

Abcam Scientific Support

Answered on Apr 17 2014

Question

What is the limit of detection for the Caspase-1 Assay (ab39412)?

Read More

Abcam community

Verified customer

Asked on Oct 30 2013

Answer


ab39412 has sensitivity of 0.05 - 0.5 ng.

Read More

Kevin Hanson

Abcam Scientific Support

Answered on Oct 30 2013

Question

Kindly contacted us regarding the Cell Lysis Buffer included in this kit. They are using a virus in there samples and are worried whether the detergent in the buffer may inactivate this virus. They ask if we might be able to share which detergent(s) are used in this lysis buffer or if they might use an alternative buffer such as RIPA?

Read More

Abcam community

Verified customer

Asked on Aug 13 2013

Answer

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.

Read More

Keith B

Abcam Scientific Support

Answered on Aug 13 2013

Question

Customer is getting a very high reading in her sample buffer only (blank) control: Empty reads about 300, blank about 800. Is this normal?
Can rule out bleed over from other wells as well as a contamination with the standard.

Read More

Abcam community

Verified customer

Asked on Dec 04 2012

Answer

Vielen Dank für Ihren Anruf.

Ist es vielleicht möglich, uns eine Excel Tabelle mit Ihren Ergebnissen zukommen zu lasen? Das Labor würde sich gerne einmal Ihre Messungen ansehen.

Ich freue mich auf Ihre Antwort.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

Read More

Abcam Scientific Support

Answered on Dec 04 2012

Question

What is the range of sensitivity of this assay?

Read More

Abcam community

Verified customer

Asked on Oct 11 2011

Answer

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 you have any more questions. 

Read More

Abcam Scientific Support

Answered on Oct 11 2011

Question

I'm using mouse left ventricle with this kit. I was wondering what type of fold increase has been seen before when testing this kit?

Read More

Abcam community

Verified customer

Asked on Sep 13 2011

Answer

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 activation after apoptosis induction. We do have the following citations for this kit which will be of help to you to give you an idea: Caspase-1 Fluorometric Assay Kit Cheng, S. C. et. al. The dectin-1/inflammasome pathway is responsible for the induction of protective T-helper 17 responses that discriminate between yeasts and hyphae of Candida albicans. J. Leukoc. Biol., Aug 2011; 90: 357 - 366. Cihakova D et al (2008) Am. J. Pathol. 172: 1195-1208. Coletti, D., et al. (2002) EMBO J. 21:631-642. Duncan JA et al (2009) J. Immunol.; 182: 6460-6469Koenen, T. b. et. al. Hyperglycemia Activates Caspase-1 and TXNIP-Mediated IL-1β Transcription in Human Adipose Tissue. Diabetes, Feb 2011; 60: 517 - 524.  I hope this information helps. Please contact us with any other questions.

Read More

Abcam Scientific Support

Answered on Sep 13 2011

Question

How should tissue be prepared to use with the kit? Any specific protocol? After lysing tissue in the cell lysis buffer, can I use this tissue lysate in a Western blot?

Read More

Abcam community

Verified customer

Asked on Sep 01 2011

Answer

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 Step 4 on the protocol. 2) You can use the lysate for Western blot. I hope this information is helpful, but please let me know if you have any further questions.

Read More

Abcam Scientific Support

Answered on Sep 01 2011

Question

Products ab39470 and ab39412. What sensitivity (pg, ng, ug?) do these products range from, for the detection of Caspase1?

Read More

Abcam community

Verified customer

Asked on Dec 16 2009

Answer

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 contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on Dec 17 2009

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