While fluorimetric assay
kits for the detection and inhibitor screening of Tumor Necrosis Factor alpha
(TNF-α) Converting Enzyme (TACE) currently exist, none of these have been
able to take full advantage of the performance provided by long
wavelength FRET technologies. Until now.
AnaSpec is pleased to
introduce the SensoLyteTM 520 TACE Assay Kit, offering fluorescence
detection at the industry’s longest wavelengths. With superior fluorescence
quantum yield and longer emission wavelength, this kit shows less autofluorescence
interference from test compounds and cellular components, and provides better
assay sensitivity than comparable kits using shorter-wavelength FRET substrates.
The SensoLyte™ 520
TACE Activity Assay Kit uses 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 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-α) Converting Enzyme (TACE),
also called ADAM17 or α-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.1 TACE, the protease responsible for the generation of mature TNF-α, plays an important role in acute and chronic inflammation. Since TNF-α is a crucial mediator in the inflammatory process, considerable efforts have been made in the research and development of anti-TNF-α agents, for the purpose of reducing the severity of inflammatory responses in disease states.2,3 The inhibition
of TACE by a pharmacological agent may represent an alternative approach to
modulate the effect of TNF-α.4 TACE is also responsible for the
proteolytic cleavage of amyloid precursor protein, L-selectin, transforming
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AnaSpec’s Detection Reagents and Kits catalog
1. Peschon, JJ. et al. Science 282, 1281 (1998).
2. Feldman, M. et al. Transplant Proc. 30, 4126 (1998).
3. Siegel, SA. et al. Cytokine 7, 15 (1995)
4. Levin, JI. et al. Bioorg. Med. Chem. Lett. 13, 2799 (2003).
5. Buxbaum, JD. et al. J. Biol. Chem. 273, 27765 (1998).
6. Smalley, DM. and Ley, K. J. Cell. Mol. Med. 9, 255 (2005).