SensoLyte® 520 Calpain Activity Assay Kit

Catalog No. ABIN1882423

Quick Overview for SensoLyte® 520 Calpain Activity Assay Kit (ABIN1882423)

Target: Calpain

Detection Method: Fluorometric

Application: Activation (Act), Fluorescence Resonance Energy Transfer Microscopy (FRET)

Product Details

Brand: SensoLyte®

Characteristics: 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.

Application Details

Comment:

FRET-based Assay Kit

Restrictions: For Research Use only

Handling

Handling Advice: Aliquot as needed to avoid multiple freeze-thaw cycles. Protect Components A and B from light and moisture.

Storage: -80 °C

Storage Comment: Store all kit components, except Component D, at -20 °C. Store Component D at -80 °C. Component C can be stored at room temperature for convenience.

References

Jock, Leggett, Schuler, Callaghan, Levin: "Effect of partial bladder outlet obstruction and reversal on rabbit bladder physiology and biochemistry: duration of recovery period and severity of function." in: BJU international, Vol. 114, Issue 6, pp. 946-54, (2014) (PubMed).

Kawashima, Umehara, Noma, Kawai, Shitanaka, Richards, Shimada: "Targeted disruption of Nrg1 in granulosa cells alters the temporal progression of oocyte maturation." in: Molecular endocrinology (Baltimore, Md.), Vol. 28, Issue 5, pp. 706-21, (2014) (PubMed).

Callaghan, Johnson, Neumann, Leggett, Schuler, Levin: "The effect of partial outlet obstruction on calpain and phospholipase-2 activities: analyzed by severity and duration." in: Molecular and cellular biochemistry, Vol. 381, Issue 1-2, pp. 217-20, (2013) (PubMed).

Kawashima, Liu, Mullany, Mihara, Richards, Shimada: "EGF-like factors induce expansion of the cumulus cell-oocyte complexes by activating calpain-mediated cell movement." in: Endocrinology, Vol. 153, Issue 8, pp. 3949-59, (2012) (PubMed).

Novgorodov, Chudakova, Wheeler, Bielawski, Kindy, Obeid, Gudz: "Developmentally regulated ceramide synthase 6 increases mitochondrial Ca2+ loading capacity and promotes apoptosis." in: The Journal of biological chemistry, Vol. 286, Issue 6, pp. 4644-58, (2011) (PubMed).

Target Details

Target: Calpain

Background: 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.