This week AnaSpec introduced the SensoLyte™ 520 TACE Assay Kit, offering fluorescence detection at the wavelengths. With higher fluorescence quantum yield and longer emission wavelength, this kit shows less autofluorescence interference from test compounds and cellular components, and can provide assay sensitivity than comparable kits using shorter-wavelength FRET substrates.
The SensoLyte™ 520 TACE Activity Assay Kit uses a FRET peptide substrate for the continuous measurement of enzyme activity. The FRET pair is composed of 5-FAM (5-carboxyfluorescein) and QXL™ 520, one of AnaSpec’s proprietary fluorescence quenchers. In the intact FRET peptide, the fluorescence of 5-FAM is quenched by QXL™ 520.
Upon cleavage of the FRET peptide by the active enzyme, the fluorescence of 5-FAM is recovered, and can be continuously monitored at excitation/emission = 490 nm/520 nm. Packaged in a 96-well format, the kit can be used to detect the activity of TACE or screen for TACE inhibitors.
Tumor Necrosis Factor alpha (TNF-a) Converting Enzyme (TACE), also called ADAM17 or ß-Secretase belongs to the ADAM (A Disintegrin and Metalloprotease) family of proteins involved in myogenesis, neurogenesis, fertilization through the ectodomain shedding of cell surface proteins.
TACE plays a crucial role in acute and chronic inflammation. Since it is a crucial mediator in the inflammatory process, considerable efforts have been made in the research and development of anti-TNF-?a agents, for the purpose of reducing the severity of inflammatory responses in disease states.
The inhibition of TACE by a pharmacological agent may represent an alternative approach to modulate the effect of TNF-a. ?TACE is also responsible for the proteolytic cleavage of amyloid precursor protein, L-selectin, transforming growth factor-a.