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Protease assays are widely used for the investigation of protease inhibitors and detection of protease activities.

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Functional Studies - Protease Activity Assay Kit (Fluorometric - Red) (AB112153), expandable thumbnail

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Key facts

Detection method

Fluorescent

Sample types

Plasma, Cell culture extracts, Other biological fluids

Assay type

Enzyme activity

Assay time

1h

Associated Products

Select an associated product type

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What's included?

500 Test
Components
Assay Buffer
1 x 30 mL
Protease Substrate
1 x 300 µL
Trypsin
1 x 100 µL

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Protease assays are widely used for the investigation of protease inhibitors and detection of protease activities.

Key facts

Detection method

Fluorescent

Sample types

Plasma, Cell culture extracts, Other biological fluids

Assay type

Enzyme activity

Assay time

1h

Assay Platform

Microplate reader

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

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. ab112153 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. ab112153 uses a red fluorescent casein conjugate that 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 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. Its signal can be easily read at Ex/Em = 540 /590 nm. ab112153 has been used for screening protease inhibitors in a HTS mode. Visit our for tips and troubleshooting.

Supplementary info

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

Activity summary

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.

Associated diseases and disorders

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.

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  • Functional Studies - Protease Activity Assay Kit (Fluorometric - Red) (ab112153), expandable thumbnail

    Functional Studies - Protease Activity Assay Kit (Fluorometric - Red) (ab112153)

    Trypsin protease activity was analyzed by using ab112153. Protease substrate was incubated with 3 units of 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 fluorescence microplate reader with a filter set of Ex/Em = 540/590 nm. Samples were done in triplicate.

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Product protocols

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