Assay-kits

SensoLyte® 520 Calpain Activity Assay Kit Fluorimetric - 1 kit

$552.00
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  • Cat.Number : AS-72149
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    In stock
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The calpains are a family of intracellular Ca2+ dependent cysteine proteases. The best-characterized calpains are isoforms 1 (µ) and 2 (m). They respond to Ca2+ signals by cleaving many specific proteins, thereby irreversibly modifying their function(s). Calpains are implicated in a variety of Ca2+ regulated cellular processes as well as various pathological phenomena such as ischemic injury, muscular dystrophy, diabetes, cataract, atherosclerosis, Alzheimer’s disease, and cancer. Calpains represent potential therapeutic targets for drug discovery.
The SensoLyte® 520 Calpain Assay Kit is optimized for detecting calpain activity. This kit contains a novel internally quenched 5-FAM/QXL™ 520 FRET substrate. Calpain protease cleaves the FRET substrate into two separate fragments resulting in the release of 5-FAM fluorescence which can be monitored at excitation/emission= 490/520 nm. Increase in fluorescence is proportional to the calpain activity. The assay can detect both calpain 1 (µ) and 2 (m) activities and is ideal for kinetic study of these enzymes. The long wavelength fluorescence of 5-FAM is less interfered by the autofluorescence of components in biological samples and test compounds. The assays are performed in a convenient 96-well microplate format.

Specifications

Packaging
Kits components
  • Component A: 5-FAM/QXL 520 Calpain Substrate, Ex/Em=490 nm/520 nm upon cleavage: 1 mM, 50 µL Component B: 5-FAM, Fluorescence Reference Standard, Ex/Em=490 nm/520 nm: 1 mM, 10 µL Component C: Assay Buffer: 20 mL Component D: Human Calpain: 0.5 mg/mL, 100 µL Component E: Calpain Inhibitor: 50 µM, 10 µL Component F: DTT: 1 M, 20 µL
Chemistry
UniProt number
  • P07384
Properties
Absorbance (nm)
  • 490
Emission (nm)
  • 520
Storage & stability
Storage Conditions
  • Store component D at -80°C. Store all other components at -20°C. Component C can be stored at room temperature for convenience. Protect components A and B from light and moisture.
Activity
Application
Biomarker Target
Detection Method
Detection Limit
  • 32.5 ng/ml
Research Area
Sub-category Research Area
Usage
  • Research use

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Citations

Effect of partial outlet obstruction and reversal on rabbit bladder physiology and biochemistry: Duration of recovery and severity of function

BJU Int. . 2014 Jul 15 ; 114(6) 946 | DOI : 10.1111/bju.12687

  • M. Jock

Targeted disruption of Nrg1 in granulosa cells alters the temporal progression of oocyte maturation

Mol Endocrinol. . 2014 Mar 20 ; 28(5) 706 | DOI : 10.1210/me.2013-1316

  • I. Kawashima

The effect of partial outlet obstruction on calpain and phospholipase-2 activities: analyzed by severity and duration

Mol Cell Biochem . 2013 Jun 05 ; 381(1-2) 217 | DOI : 10.1007/s11010-013-1705-8

  • C.M. Callaghan

EGF-Like factors induce expansion of the cumulus cell-oocyte complexes by activating calpain-mediated cell movement

Endocrinology . 2012 Jun 06 ; 153(8) 3949 | DOI : 10.1210/en.2012-1059

  • I. Kawashima

Developmentally Regulated Ceramide Synthase 6 Increases Mitochondrial Ca2+ Loading Capacity and Promotes Apoptosis

J Biol Chem. . 2010 Dec 10 ; 286(6) 4644 | DOI : 10.1074/jbc.M110.164392

  • S. Novgorodov

Radiocontrast Agent Diatrizoic Acid Induces Mitophagy and Oxidative Stress via Calcium Dysregulation

Int. J. Mol. Sci. . 2019 Aug 21 ; 20(17) 4074 | DOI : https://doi.org/10.3390/ijms20174074

  • DB. Ward
  • et al

Capsaicin Synergizes with Camptothecin to Induce Increased Apoptosis in Human Small Cell Lung Cancers via the Calpain Pathway

Biochem Pharmacol. . 2017 Apr 01 ; 129 54 | DOI : 10.1016/j.bcp.2017.01.004

  • JR. Friedman
  • et al