AnaSpec has introduced the SensoLyte 520 TACE Assay Kit, offering fluorescence detection at the industry's longest wavelengths
With higher fluorescence quantum yield and longer emission wavelength, the kit shows less autofluorescence interference from test compounds and cellular components, and provides greater 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 = 490nm/520nm.
Packaged in a convenient 96-well format, the kit can be used to detect the activity of TACE or screen for TACE inhibitors.
Tumor Necrosis Factor alpha (TNF- alpha) Converting Enzyme (TACE), also called ADAM17 or b-Secretase belongs to the ADAM (A Disintegrin and Metalloprotease) family of proteins involved in myogenesis, neurogenesis, fertilisation through the ectodomain shedding of cell surface proteins (Ref 1).
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 (Refs 2,3).
The inhibition of TACE by a pharmacological agent may represent an alternative approach to modulate the effect of TNF- alpha (Ref 4).
TACE is also responsible for the proteolytic cleavage of amyloid precursor protein, L-selectin, transforming growth factor-alpha (Refs 1,5,6).
Ref 1 - Peschon, JJ et al, Science 282, 1281 (1998).
Ref 2 - Feldman, M et al, Transplant Proc 30, 4126 (1998).
Ref 3 - Siegel, SA.
et al, Cytokine 7, 15 (1995) Ref 4 - Levin, JI et al, Bioorg.
Med Chem Lett 13, 2799 (2003).
Ref 5 - Buxbaum, JD et al, J Biol Chem 273, 27765 (1998).
Ref 6 - Smalley, DM and Ley, K J Cell Mol Med 9, 255 (2005).
