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AnaSpec, EGT Group is pleased to introduce a new long wavelength fluorimetric assay kit for determining Dipeptidyl Peptidase IV (DPPIV) activity, the SensoLyte® Rh110 DPPIV Assay Kit. This kit contains a substrate that emits in the green range of 520 nm when cleaved by DPPIV. The kit provides a convenient assay for enzyme inhibitors and activators screening or for the continuous assay of enzyme activity. Cleavage of the fluorogenic substrate by DPPIV generates the Rh110 (rhodamine 110) fluorophore which has a bright green fluorescence. Fluorescence can be monitored at excitation/emission=490/520 nm. The longer wavelength spectra and higher extinction coefficient of Rh110 provide greater sensitivity and less interference from other reaction components.
Dipeptidyl Peptidase IV (DPPIV, DPP-4, CD26) is a cell surface serine protease, which cleaves Xaa-Pro or less efficiently Xaa-Ala dipeptides from the N-terminus.1-3 DPPIV possesses an identical function to the T-cell surface antigen (CD26).1 DPPIV is commonly found in mammalian tissues and is highly expressed in hepatocytes, pancreatic epithelial cells, intestinal epithelium, and kidney cortex.3 DPPIV plays an important role in glucose homeostasis by proteolysis of incretins such as glucagon-like peptide-1 (GLP-1). Due to its role in degradation of GLP-1, DPPIV is a therapeutic target for type 2 diabetes.
The SensoLyte®AMC DPPIV Assay Kit, which employs an AMC fluorogenic substrate, is also available from AnaSpec. This kit is optimized for screening of enzyme inhibitors. This kit contains an AMC labeled substrate that has a high reactivity and low background. Upon protease cleavage, this substrate generates the AMC fluorophore emitting bright blue fluorescence that can be monitored at excitation/emission=354/442 nm. Increase in AMC fluorescence is proportional to DPPIV activity.
References
1. Zhu, L. et al, J. Biol. Chem. 278, 22418 (2003).
2. Mentlein, R. Regulatory Peptides 85, 9 (1998).
3. Qi, SY. et al, Biochem. J. 373, 179 (2003).
AnaSpec, EGT Group
AnaSpec, EGT Group (anaspec.com) is a leading provider of integrated proteomics and genomics solutions™ for worldwide life science research. AnaSpec offers expertise in peptides, detection reagents, antibodies, assay kits, amino acids, 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, fluorescent dyes, and oligonucleotide synthesis. AnaSpec is certified to ISO 9001:2008.
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