This kit employs a natural substrate, elastin that is labeled with the 5-FAM fluorophore and the QXL™ 520 quencher. Proteolytic cleavage of labeled elastin separates the donor from the quencher and yields brightly green fluorescence, which can be continuously monitored at excitation/emission (Ex/Em) = 488 nm/520 nm. Increase in fluorescence intensity is directly proportional to elastase activity. The kit can be used to continuously measure the kinetics of elastase activity. As well, this highly sensitive kit is homogeneous – it does not require any separation steps.
Elastases catalyze the hydrolysis of elastin, the chief constituent of elastic fibers that together with collagen determines the mechanical properties of connective tissue. Elastases have many other natural substrates these include proteoglycans, collagens and fibronectin.1-4 Implicated in many diseases such as pulmonary emphysema, cystic fibrosis, infections, inflammation and atherosclerosis, elastases are considered important drug targets.5
AnaSpec, Eurogentec Group also provides another green emitting elastase assay, the SensoLyte® Rh110 Elastase Assay Kit. This assay uses a substrate that is labeled with a fluorogenic dye called Rh110 (rhodamine 110). Upon cleavage by elastase, the substrate generates free Rh110 fluorophore, which has a bright green fluorescence. Signal can be detected at Ex/Em =490/520 nm. The longer wavelength spectra and higher extinction coefficient of this kit and the SensoLyte® Green Elastase kits provide greater sensitivity and less interference from other reaction components.
1. Kafienah, W. et al. Biochem J. 330, 897 (1998).
2. Carlo, L. et al. J. Biol. Chem. 255, 12006 (1980).
3. Mainardi, CL. et al. J. Biol. Chem. 255, 5435 (1980).
4. McDonald, JA. et al. J. Biol. Chem. 255, 8848 (1980).
5. Bieth, JG. J. Soc. Biol. 195, 173 (2001).
About AnaSpec, Eurogentec Group
AnaSpec (anaspec.com), Eurogentec Group is a leading provider of integrated proteomics and genomics solutionsTM for worldwide life science research. As a subsidiary of Eurogentec, AnaSpec offers expertise in peptides, detection reagents, antibodies, assay kits, oligonucleotides, and qPCRs. AnaSpec carries a broad product line of biochemicals and reagents for basic research, high-throughput screening and drug discovery. AnaSpec also provides premier custom services including peptide synthesis, antibody production, assay development and oligonucleotide synthesis. AnaSpec is certified to ISO 9001:2008.