Recombinant Human ADAMTS4 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human ADAMTS4 Protein, CF Summary

Details of Functionality
Measured by its ability to cleave the fluorogenic peptide substrate, Abz-TEGEARGSVI-Dap(Dnp)-KK-NH2. The specific activity is >7 pmol/min/μg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived human ADAMTS4 protein
Phe213-Cys685, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Phe213
Structure / Form
Recombinant Human ADAMTS4 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
Protein/Peptide Type
Recombinant Enzymes
Gene
ADAMTS4
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
53 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
58 kDa, reducing conditions
Publications
Read Publications using
4307-AD in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Sodium Acetate, CaCl2 and NaCl.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 50 mM HEPES, 50 mM NaCl, 1 mM CaCl2, 0.05% Brij-35, pH 7.5
  • Recombinant Human ADAMTS4 (Catalog # 4307-AD)
  • Substrate: WAAG-3R (Anaspec, Catalog # 60431-1), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhADAMTS4 to 10 µg/mL in Assay Buffer.
  2. Dilute Substrate to 50 µM in Assay Buffer.
  3. Load 50 µL of 10 µg/mL of rhADAMTS4 into a plate, and start the reaction by adding 50 µL of 50 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
  4. Read at excitation and emission wavelengths of 340 nm and 420 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     

     *Adjusted for Substrate Blank
      **Derived using calibration standard Abz-Gly-OH (Bachem, Catalog # E-2920).

Per Well:
  • rhADAMTS4: 0.5 µg
  • Substrate: 25 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human ADAMTS4 Protein, CF

  • a disintegrin-like and metalloprotease (reprolysin type) with thrombospondintype 1 motif, 4
  • ADAM metallopeptidase with thrombospondin type 1 motif, 4
  • ADAM-TS 4
  • ADAMTS2
  • ADAMTS-2
  • ADAMTS4
  • ADAM-TS4
  • ADAMTS-4
  • ADMP-1
  • ADMP-1EC 3.4.24.82
  • Aggrecanase 1
  • aggrecanase-1
  • EC 3.4.24
  • KIAA0688A disintegrin and metalloproteinase with thrombospondin motifs 4

Background

ADAMTS4 (a disintegrin and metalloproteinase with thrombospondin motifs 4), also known as aggrecanase-1, is a member of the family of secreted zinc proteases with a multi-domain structure (1-3). The protein precursors consist of a signal peptide and the following domains: pro, catalytic, disintegrin-like, TS type 1 motif, cysteine-rich, and spacer. It is the only ADAMTS identified that has one TS type I motif. It is an active protease effectively cleaving alpha -2-macroglobulin and aggrecan at multiple sites, and is inhibited by TIMP-3 with inhibition constants in subnanomolar range (4-6). It receives great attention due to the elevation in its mRNA level after treatment with Interleukin-1 (7). However, in a mouse model of osteoarthritis, ADAMTS4 knock-out mice did not exhibit any significant protective effect (8). The purified rhADAMTS4 starts at the catalytic domain and ends before the spacer domain. If desired, the aggrecanase activity can be inhibited by 5 mM 1 10‑phenanthroline and Recombinant Human TIMP‑3 (Catalog # 973-TM).

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Publications for ADAMTS4 (4307-AD)(6)

We have publications tested in 4 confirmed species: Human, Mouse, Rat, N/A.

We have publications tested in 5 applications: Bioassay, EnzAct, Enzyme Assay, In Vivo, WB Ctrl.


Filter By Application
Bioassay
(2)
EnzAct
(1)
Enzyme Assay
(1)
In Vivo
(1)
WB Ctrl
(1)
All Applications
Filter By Species
Human
(1)
Mouse
(2)
Rat
(1)
N/A
(2)
All Species
Showing Publications 1 - 6 of 6.
Publications using 4307-AD Applications Species
T Fontanil, S ?lvarez-Te, M? Villaronga, Y Mohamedi, L Solares, A Moncada-Pa, JA Vega, O Garc¡a-Su , M P‚rez-Bast, JM Garc¡a-Ped, AJ Obaya, S Cal Cleavage of Fibulin-2 by the aggrecanases ADAMTS-4 and ADAMTS-5 contributes to the tumorigenic potential of breast cancer cells Oncotarget, 2017;0(0):. 2017 [PMID: 28099917] (Enzyme Assay, Human) Enzyme Assay Human
Lemarchant S, Pomeshchik Y, Kidin I, Karkkainen V, Valonen P, Lehtonen S, Goldsteins G, Malm T, Kanninen K, Koistinaho J ADAMTS-4 promotes neurodegeneration in a mouse model of amyotrophic lateral sclerosis. Mol Neurodegener, 2016;11(1):10. 2016 [PMID: 26809777] (Bioassay, Mouse) Bioassay Mouse
Rao N, Ke Z, Liu H, Ho C, Kumar S, Xiang W, Zhu Y, Ge R ADAMTS4 and its proteolytic fragments differentially affect melanoma growth and angiogenesis in mice. Int J Cancer, 2013;133(2):294-306. 2013 [PMID: 23319426] (WB Ctrl, N/A) WB Ctrl N/A
Devel L, Beau F, Amoura M, Vera L, Cassar-Lajeunesse E, Garcia S, Czarny B, Stura E, Dive V Simple pseudo-dipeptides with a P2&#039; glutamate: a novel inhibitor family of matrix metalloproteases and other metzincins. J Biol Chem, 2012;287(32):26647-56. 2012 [PMID: 22689580] (EnzAct, N/A) EnzAct N/A
Tauchi R, Imagama S, Natori T The endogenous proteoglycan-degrading enzyme ADAMTS-4 promotes functional recovery after spinal cord injury. J Neuroinflammation, 2012;9(0):53. 2012 [PMID: 22420304] (In Vivo, Rat) In Vivo Rat
Weaver MS, Workman G, Cardo-Vila M, Arap W, Pasqualini R, Sage EH Processing of the matricellular protein hevin in mouse brain is dependent on ADAMTS4. J. Biol. Chem., 2010;285(8):5868-77. 2010 [PMID: 20018883] (Bioassay, Mouse) Bioassay Mouse

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Bioinformatics

Gene Symbol ADAMTS4
Uniprot