WAAG - 3R, Aggrecanase (ADAM - TS - 4) FRET Substrate
WAAG - 3R, Aggrecanase (ADAM - TS - 4) FRET Substrate
Product Name: WAAG-3R, Aggrecanase (ADAM-TS-4) FRET Substrate
Catalog Number: AS-60431-1 (1 mg) Lot Number: See label on vial
Sequence: Abz-Thr-Glu-Gly-Glu-Ala-Arg-Gly-Ser-Val-Ile-Dap(Dnp)-Lys-Lys-NH2
(3-letter code)
Abz-TEGEARGSVI-Dap(Dnp)-KK-NH2 (1-letter code)
Molecular Weight: 1645.8
% Peak Area by HPLC: ≥ 95
Appearance: Lyophilized yellow powder
Peptide Reconstitution: WAAG-3R peptide is freely soluble in H2O.
Storage: WAAG-3R peptide is shipped at ambient temperature. Upon receipt, store lyophilized peptide
at –20°C or lower. Reconstituted peptide can be aliquoted and stored at –20°C or lower.
Description: This FRET peptide was used in an ADAM-TS-4 (Aggrecanase-1) assay. Ex/Em =
340/420 nm. Ref: Zhang, Y. et al. J. Pharmacol. Exp. Ther. 309, 348 (2004).
Additional Information: Listed below are relevant information that may provide a guideline on how to
use this product. End users will have to adapt to their own specific applications.
The ADAM-TS-4 (Aggrecanase-1) assay was performed using a fluorescent peptide Abz-
TEGEARGSVI-Dap(Dnp)-KK (denoted as WAAG-3R, custom synthesized by AnaSpec, Inc.). The
assay buffer contains 50 mM HEPES, pH 7.5, 100 mM NaCl, 5 mM CaCl2, 0.1% CHAPS, and 5%
glycerol. Total reaction volume is 100 μl. The recombinant Agg-1 proteins generated at Wyeth
Research (final concentration of 5 μg/ml in the assay) were pretreated with the various concentrations
of the compound for 10 to 15 min at 37°C. The reaction was initiated by addition of the WAAG-3R
substrate at a final concentration of 25 μM- Zhang, Y. et al. J. Pharmacol. Exp. Ther. 309, 348 (2004).
The peptide sequence was Abz-TEGEARGSVI-Dap(Dnp)-KK (Anaspec) and derived from the
aggrecanase cleavage site of aggrecan. The final concentration of substrate in the assay was μ25 M.
The buffer used in this assay was 50 mM HEPES (pH 7.5), 100 mM NaCl, 5 mM CaCl2, 0.1% CHAPS,
and 5% glycerol. Compounds (in duplicate) were serially diluted from 2 mM to 0.01 μM in 100%
DMSO. The total reaction volume was 100 μL. The buffer and enzyme were added first followed by
addition of 10× inhibitor. Enzyme and buffer alone samples were included in order to obtain the
maximal rate of substrate cleavage. The reaction was allowed to stand for 15 min at 30°C- Mosyak, L.
et al. Protein Science 17, 16 (2008).
Published Citations:
Zhang, Y. et al. J. Pharmacol. Exp. Ther. 309, 348 (2004).
Mosyak, L. et al. Protein Science 17, 16 (2008).
Related Products:
Name Cat # SSSize
SensoLyte® 520 Aggrecanase-1 Assay Kit *Fluorimetric* AS-72114 1 kit
For Research Use Only
