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AB112152

Protease Activity Assay Kit (Fluorometric - Green)

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(3 Publications)

Protease Activity Assay Kit ab112152 uses a fluorescent casein conjugate as a generic substrate for a broad spectrum of proteases. Cleavage by proteases relieves quenching of the fluorescent dye, with fluorometric readout (Ex/Em 490/525 nm).
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Functional Studies - Protease Activity Assay Kit (Fluorometric - Green) (AB112152)
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Supplier Data

Functional Studies - Protease Activity Assay Kit (Fluorometric - Green) (AB112152)

Trypsin protease activity was analyzed by using ab112152. Protease substrate was incubated with 1 unit trypsin in the kit assay buffer. The control wells had protease substrate only (without trypsin). The fluorescence signal was measured starting from time 0 when trypsin was added using a microplate reader with a filter set of Ex/Em = 490/525 nm. Samples were done in triplicates.

Key facts

Detection method

Fluorescent

Sample types

Cell culture extracts, Other biological fluids, Plasma, Serum

Assay type

Enzyme activity

Assay time

1h

Assay Platform

Microplate reader

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Enzyme activity assay": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Protease assays are widely used for the investigation of protease inhibitors and detection of protease activities. Monitoring various protease activities has become a routine task for many biological laboratories. Some proteases have been identified as good drug development targets.

ab112152 Protease Activity Assay Kit is an ideal choice to perform routine assays for the isolation of proteases, or for identifying the presence of contaminating proteases in protein samples. ab112152 uses a fluorescent casein conjugate which is proven to be a generic substrate for a broad spectrum of proteases (e.g. trypsin, chymotrypsin, thermolysin, proteinase K, protease XIV, and elastase). In the intact substrate, casein is heavily labeled with a green fluorescent dye, resulting in significant fluorescence quenching. Protease-catalyzed hydrolysis relieves its quenching effect, yielding brightly fluorescent dye-labeled short peptides. The increase in fluorescence intensity is directly proportional to protease activity. The assay can be performed in a convenient 96-well or 384-well microtiter plate format and readily adapted to automation. Its signal can be easily read with a fluorescence microplate reader at Ex/Em = 490/525 nm using FITC filter set.

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What's included?

{ "values": { "500Test": { "sellingSize": "500 Test", "publicAssetCode":"ab112152-500Test", "assetComponentDetails": [ { "size":"1 x 30 mL", "name":"2X Assay Buffer", "number":"AB112152-CMP01", "productcode":"" }, { "size":"1 x 300 µL", "name":"Protease Substrate", "number":"AB112152-CMP03", "productcode":"" }, { "size":"1 x 100 µL", "name":"Trypsin", "number":"AB112152-CMP02", "productcode":"" } ] } } }

Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
Multi
Storage information
Please refer to protocols

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Proteases also known as peptidases or proteinases are enzymes that catalyze the breakdown of proteins into smaller peptides or amino acids by cleaving peptide bonds. These enzymes vary in mass typically ranging from 20 to 90 kDa depending on the specific type and its functional domain. They are expressed across different tissues and cellular compartments with high concentrations in digestive organs lysosomes and blood plasma. The mechanical process of protease activity is central to many physiological processes making their precise function an important focus of study in biology.
Biological function summary

Proteases play critical roles in maintaining cellular functions including protein catabolism cell signaling and the regulation of the cell cycle. They are often components of large complexes such as the proteasome where they help degrade ubiquitinated proteins ensuring protein homeostasis. Proteases also assist in activating precursor molecules into biologically active forms involving themselves in processes like blood coagulation immune responses and apoptosis. Their activity is essential in remodeling extracellular matrix and processing bioactive molecules further influencing diverse physiological pathways.

Pathways

Proteases significantly impact both the proteolytic and the ubiquitin-proteasome pathways. Within the proteolytic pathway enzymes break down proteins into peptides and amino acids for cellular recycling and energy production. The ubiquitin-proteasome pathway involving ubiquitin-related proteins is important for regulating protein degradation and turnover impacting cellular functions and stress responses. Proteases also interact with other proteins such as kinases and phosphatases which facilitate cellular signaling cascades reflecting their participation in broader biological networks.

Many proteases have associations with cancer and neurodegenerative diseases. Aberrant protease activity can lead to uncontrolled cell proliferation or faulty cell death contributing to carcinogenesis. In neurodegenerative disorders such as Alzheimer's disease improper protease activity results in the accumulation of misfolded proteins like amyloid-beta peptides disrupting neural function. Proteases also have connections with proteins such as tau and APP in Alzheimer's disease indicating their complex role in pathogenesis and providing potential targets for therapeutic interventions.

Product protocols

Target data

Publications (3)

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BMC veterinary research 20:368 PubMed39152418

2024

Effects of dietary supplementation with Bacillus velezensis on the growth performance, body composition, antioxidant, immune-related gene expression, and histology of Pacific white shrimp, Litopenaeus vannamei.

Applications

Unspecified application

Species

Unspecified reactive species

Arwa E M Abdelsamad,Rashad E M Said,Mona Assas,Alkhateib Y Gaafar,Awatef H Hamouda,Aldoushy Mahdy

The ISME journal 16:1970-1979 PubMed35545659

2022

Bacterial matrix metalloproteases and serine proteases contribute to the extra-host inactivation of enteroviruses in lake water.

Applications

Unspecified application

Species

Unspecified reactive species

Marie-Hélène Corre,Virginie Bachmann,Tamar Kohn

Aging cell 17:e12710 PubMed29730901

2018

HTRA1, an age-related macular degeneration protease, processes extracellular matrix proteins EFEMP1 and TSP1.

Applications

Unspecified application

Species

Unspecified reactive species

Michael K Lin,Jin Yang,Chun Wei Hsu,Anuradha Gore,Alexander G Bassuk,Lewis M Brown,Ryan Colligan,Jesse D Sengillo,Vinit B Mahajan,Stephen H Tsang
View all publications
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