Description
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
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. Visit our for tips and troubleshooting.
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.
