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
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
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.
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 DTT 1 x 0.4ml YVAD-AFC 1 x 0.5ml -
Research areas
-
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
see all
Images
-
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)Titration of the caspase 1 (ab39901) (background signal subtracted, duplicates; +/- SD).
