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.

AnaSpec’s SensoLyte^® 520 TACE Assay Kit offers 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 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. 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 growth factor-α.^1,5,6

Looking for custom development of fluorimetric assays? Please click here or email service@anaspec.com to see how AnaSpec can help.

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References:

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 KJ. Ley, Cell. Mol. Med. 9, 255 (2005).