Assay-kits

SensoLyte® 520 Aggrecanase-1 Assay Kit Fluorimetric - 1 kit

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  • Cat.Number : AS-72114
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    In stock
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Aggrecanases belong to ADAMTS (A disintegrin and metalloprotease with thrombospondin motif) family of proteases. Aggrecanases cleave aggrecan, the major structural component of cartilage. Aggrecanase-1 (ADAMTS-4) is a major aggrecanase in human osteoarthritic cartilage.
The SensoLyte® 520 Aggrecanase-1 Assay Kit uses 5-FAM as the fluorophore and TAMRA as the quencher in the FRET peptide substrate. This substrate is used for continuous measurement of the enzyme activity. In an intact FRET peptide, the fluorescence of 5-FAM is quenched by TAMRA. Upon the cleavage of the FRET peptide by ADAMTS-4, the fluorescence of 5-FAM is recovered, and can be continuously monitored at excitation/emission = 490 nm/520 nm. Increase in FAM fluorescence is correlated to aggrecanase-1 activity. 5-FAM/TAMRA based FRET substrate is less interfered by autofluorescence of test compounds due to its longer emission wavelength. The kit can be used to detect the activity of aggrecanase-1 or screen for aggrecanase inhibitors. The assays are performed in a convenient 96-well format.

Specifications

Packaging
Kits components
  • Component A: 5-FAM/5-TAMRA, Aggrecanase-1 substrate, Ex/Em=490nm/520 nm: 60 µL Component B: 5-FAM, fluorescence reference standard, Ex/Em=490nm/520nm: 1mM, 10 µL Component C: Assay Buffer: 20 mL Component D: Control Inhibitor: 100 µM, 10 µL Component E: Stop Solution: 10 mL
Chemistry
UniProt number
  • O75173
Properties
Absorbance (nm)
  • 490
Emission (nm)
  • 520
Storage & stability
Storage Conditions
  • Store all kit components at -20°C. Protect Components A and B from light and from moisture. Components C and E can be stored at room temperature for convenience.
Activity
Application
Biomarker Target
Detection Method
Detection Limit
  • 0.1 ng/ml
Research Area
Sub-category Research Area
Usage
  • Research use

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Citations

tPA promotes ADAMTS-4-induced CSPG degradation, thereby enhancing neuroplasticity following spinal cord injury

Neurobiol Dis . 2014 Feb 25 ; 66 28 | DOI : 10.1016/j.nbd.2014.02.005

  • S. Lemarchant
  • et al

ADAMTS4_v1 is a splice variant of ADAMTS4 that is expressed as a protein in human synovium and cleaves aggrecan at the interglobular domain

Arthritis Rheumatol . 2013 Nov 01 ; 65 2866 | DOI : 10.1002/art.38102

  • S.D. Wainwright
  • et al

The endogenous proteoglycan-degrading enzyme ADAMTS-4 promotes functional recovery after spinal cord injury

J Neuroinflammation . 2012 Mar 15 ; 9 53 | DOI : 10.1186/1742-2094-9-53

  • R. Tauchi
  • et al

The nutraceutical flavonoid luteolin inhibits ADAMTS-4 and ADAMTS-5 aggrecanase activities

J Mol Med . 2011 Mar 02 ; 89(6) 611 | DOI : 10.1007/s00109-011-0741-7

  • A. Moncada-Pazos
  • et al

Structure-activity study on a series of α-glutamic acid scaffold based compounds as new ADAMTS inhibitors

Bioorg Med Chem Lett . 2011 Aug 01 ; 21(15) 4457 | DOI : 10.1016/j.bmcl.2011.06.009

  • L. Peng
  • et al