Development of High Throughput Protease Activity Detection Assay

Detection of protease activity is an important step in the research and application of protease. It can be used for both measuring specific desired protease activity as well as detection of non-specific protease activity in a sample. In the past, reagents used for such detection required prior separation of the protein substrate and amino-acid degradation products, usually by TCA precipitation. This resulted in a low throughput, multiple-steps and hazardous-material containing assay. To allow an improved high throughput (HTP) workflow we have developed a labeled protein substrate reagent. Our product, SAE0204, is designed to correlate its degradation with a fluorescent signal. The labeling dye selected is stable in a wide pH range (that is not supported using other dyes) and therefore is suitable for detection of proteases activity also in extreme pH conditions. In addition, it allows high signal to noise ratio that increase detection resolution and sensitivity. Therefore, we have developed an improved simple HTP assay for protease activity detection which is suitable for verity of samples condition including extreme PHs.