Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Cell culture extracts, Other biological fluids, Plasma, Whole Blood
Product nameProtease Activity Assay Kit (Fluorometric - Red)
See all Protease kits
Sample typePlasma, Cell culture extracts, Other biological fluids, Whole Blood
Assay typeEnzyme activity (quantitative)
Assay time1h 00m
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.
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Storage instructionsStore at -20°C. Please refer to protocols.
Components 500 tests Assay Buffer 1 x 30ml Protease Substrate 1 x 300µl Trypsin 1 x 100µl
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.
ab112153 has been referenced in 1 publication.
- Lee D et al. Application of a high-throughput microcrystal delivery system to serial femtosecond crystallography. J Appl Crystallogr 53:477-485 (2020). PubMed: 32280322