Calculator
Monitor Protease Contamination with Amplite™ Fluorimetric Assay Kits
In this issue
Light up Lysosomes with Cell Navigator™ Kits
Multiplex Apoptosis and Necrosis
Mulitplex Caspase Activities
Cal-520™, the Best Green Fluorescent Ca2+ Dye
The Rapid & Sensitive Detection of NAD, NADH, NADP & NADPH Using Amplite™ Assay Kits
Quantify Maleimide by Absorbance or Fluorescence
Fluo-8®, the Brightest Green Ca2+ Indicators
Quantify cAMP by Absorbance or Fluorescence
Optimized Fluorescence Labeling Solutions
Monitor Protease Contamination with Amplite™ Fluorimetric Assay Kits
Detect Thiol with Amplite™ Assay Kits
Detect Calcium by Absorbance or Fluorescence
AssayWise Letters 2012. Vol. 1(1)
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. On the other hand, many proteins are subject to protease-induced degradation, thus monitoring sample protease activities is often required in a variety of biological processes.
Our Amplite® Universal Fluorimetric Protease Activity Assay Kits are an ideal choice to perform routine assays for the isolation of proteases, or for identifying the presence of contaminants in protein samples. The kits use 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 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. For Kit 13500, the fluorescent signal can be easily read with a fluorescence microplate reader at Ex/Em = 490/525 nm using FITC filter set. For Kit 13501, the fluorescent signal can be easily read at Ex/Em = 540 /590 nm. Both Kit 13500 and 13501 have been used for screening protease inhibitors in a HTS mode.
Our Amplite® Universal Fluorimetric Protease Activity Assay Kits are an ideal choice to perform routine assays for the isolation of proteases, or for identifying the presence of contaminants in protein samples. The kits use 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 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. For Kit 13500, the fluorescent signal can be easily read with a fluorescence microplate reader at Ex/Em = 490/525 nm using FITC filter set. For Kit 13501, the fluorescent signal can be easily read at Ex/Em = 540 /590 nm. Both Kit 13500 and 13501 have been used for screening protease inhibitors in a HTS mode.
Key Features of Amplite® Protease Detection Kits:
- Convenient format, all the key assay components included.
- Optimized performance, optimal conditions for the detection of generic protease activity.
- Continuous, easily adapted to automation without a separation step.
- Convenient, formulated to have minimal hands-on time. No wash is required.
- Non-radioactive, no special requirements for waste treatment.
Left: Trypsin protease activity was analyzed by Amplite® Universal Fluorimetric Protease Activity Assay kit (Cat. #13500). Protease substrate was incubated with 1 unit of trypsin. The control wells had protease substrate only (without trypsin). The fluorescence signal was measured starting from time 0 when trypsin was added. Samples were done in triplicate. Right: Trypsin protease activity was analyzed by Amplite® Universal Fluorimetric Protease Activity Assay kits (Cat. #13501). Protease substrate was incubated 3 units of trypsin. The control wells had protease substrate only (without trypsin). The fluorescence signal was measured starting from time 0 when trypsin was added. Samples were done in triplicate.