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539125

Sigma-Aldrich

Protease Assay Kit

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About This Item

Code UNSPSC :
41116133
Nomenclature NACRES :
NA.84

Utilisation

sufficient for 200 tests

Niveau de qualité

Fabricant/nom de marque

Calbiochem®

Conditions de stockage

OK to freeze
protect from light

assay range

sensitivity: 10 ng/mL

Entrée

sample type biological sample(s)

Méthode de détection

colorimetric

Conditions d'expédition

wet ice

Température de stockage

−20°C

Description générale

A kit suitable for quantitative array of a wide variety of proteases in biological samples, utilizing FTC-casein as a substrate. Enzyme activities that have been detected with this assay kit include: chymotrypsin, elastase, plasminogen, PRONASE® Protease, subtilisin, thermolysin, and trypsin.
This Protease Assay Kit is suitable for the quantitative determination of protease activity in biological samples. This kit has been tested for its suitability in detecting a wide variety of proteases, including serine proteases (trypsin, chymotrypsin, thrombin, plasmin, elastase, subtilisin), cysteine proteases (papain, cathepsin B) and acid proteases (thermolysin, pepsin). Each kit provides sufficient reagents for 200 assays. When used as recommended, this kit is ~100-fold more sensitive than either the casein-gel assay or casein A280 assay. The Protease Assay Kit can detect ~10 ng/ml of protease when a 24 h incubation is used. The sensitivity can be enhanced by up to 3-fold by using a fluorometer (excitation 490 nm and emission 525 nm) rather than a spectrophotometer to detect the proteolytic end products.

Composants

A Fluorescent-Casein Substrate, Incubation Buffer, Assay Buffer Trypsin Standard, and a user protocol.

Avertissement

Toxicity: Multiple Toxicity Values, refer to MSDS (O)

Caractéristiques

Assay Time: 3 h

Stockage et stabilité

FTC-casein should be stored at -20°C and is stable for one year from the date of purchase. FTC-casein stored at 4°C is stable for 2-3 days. Several freeze-thaw cycles will lead to a minor increase in the background fluorescence, so following initial thawing, we suggest that this reagent be divided into 5 aliquots. FTC-casein is light sensitive and should be protected from light.

Protease positive control should be stored at -20°C. As with many proteases, trypsin is susceptible to autolysis and you may notice a reduction in trypsin activity with time, even if stored at -20°C.

Incubation buffer and assay buffer may be stored at 4°C or -20°C.

Autres remarques

Twining, S.S. 1984. Anal. Biochem.143, 30.
Wiesner, R., et al. 1982. Anal. Biochem.121, 290.

Informations légales

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
PRONASE is a registered trademark of Merck KGaA, Darmstadt, Germany

Code de la classe de stockage

10 - Combustible liquids


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S S Twining
Analytical biochemistry, 143(1), 30-34 (1984-11-15)
A simple inexpensive sensitive protease assay was developed using soluble fluorescein isothiocyanate (FITC)-labeled casein. Casein was reacted with FITC to form the fluorescein thiocarbamoyl derivative. This substrate is cleaved by trypsin, chymotrypsin, elastase, subtilisin, and thermolysin in a linear time-dependent
Shuofeng Yuan et al.
Protein & cell, 13(12), 940-953 (2022-04-07)
The emergence of SARS-CoV-2 variants of concern and repeated outbreaks of coronavirus epidemics in the past two decades emphasize the need for next-generation pan-coronaviral therapeutics. Drugging the multi-functional papain-like protease (PLpro) domain of the viral nsp3 holds promise. However, none
A new assay for proteases using fluorescent labeling of proteins.
R Wiesner et al.
Analytical biochemistry, 121(2), 290-294 (1982-04-01)
Zachary R Gergely et al.
Journal of biological research (Thessalonike, Greece), 25, 15-15 (2018-08-18)
The insect-trapping leaves of Dionaea muscipula provide a model for studying the secretory pathway of an inducible plant secretory system. The leaf glands were induced with bovine serum albumin to secrete proteases that were characterized via zymogram activity gels over

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