Recombinant Human ADAMTS4 Protein, CF

(6 citations)   
  • Purity
    >90%, by SDS-PAGE under reducing conditions and visualized by silver stain
  • Endotoxin Level
    <1.0 EU per 1 μg of the protein by the LAL method.
  • Activity
    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. See Activity Assay Protocol on www.RnDSystems.com.
  • Source
    Chinese Hamster Ovary cell line, CHO-derived Phe213-Cys685, with a C-terminal 10-His tag
  • Accession #
  • N-terminal Sequence
    Analysis
    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.
  • Predicted Molecular Mass
    53 kDa
  • SDS-PAGE
    58 kDa, reducing conditions
4307-AD
 
Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate, CaCl2 and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.
Assay Procedure
Materials
  • 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
Background: ADAMTS4

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).

  • Long Name:
    A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 4
  • Entrez Gene IDs:
    9507 (Human); 66015 (Rat)
  • Alternate Names:
    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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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Species
Applications
Sample Type
  1. Cleavage of Fibulin-2 by the aggrecanases ADAMTS-4 and ADAMTS-5 contributes to the tumorigenic potential of breast cancer cells
    Authors: 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
    Oncotarget, 2017;0(0):.
    Species: Human
    Sample Type: Protein
    Application: Enzyme Assay
  2. ADAMTS-4 promotes neurodegeneration in a mouse model of amyotrophic lateral sclerosis.
    Authors: Lemarchant S, Pomeshchik Y, Kidin I, Karkkainen V, Valonen P, Lehtonen S, Goldsteins G, Malm T, Kanninen K, Koistinaho J
    Mol Neurodegener, 2016;11(1):10.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Bioassay
  3. ADAMTS4 and its proteolytic fragments differentially affect melanoma growth and angiogenesis in mice.
    Authors: Rao N, Ke Z, Liu H, Ho C, Kumar S, Xiang W, Zhu Y, Ge R
    Int J Cancer, 2013;133(2):294-306.
    Species: N/A
    Sample Type: N/A
    Application: WB Ctrl
  4. The endogenous proteoglycan-degrading enzyme ADAMTS-4 promotes functional recovery after spinal cord injury.
    Authors: Tauchi R, Imagama S, Natori T
    J Neuroinflammation, 2012;9(0):53.
    Species: Rat
    Sample Type: In Vivo
    Application: In Vivo
  5. Simple pseudo-dipeptides with a P2' glutamate: a novel inhibitor family of matrix metalloproteases and other metzincins.
    Authors: Devel L, Beau F, Amoura M, Vera L, Cassar-Lajeunesse E, Garcia S, Czarny B, Stura E, Dive V
    J Biol Chem, 2012;287(32):26647-56.
    Species: N/A
    Sample Type: Fluorogenic Peptide Substrate
    Application: EnzAct
  6. Processing of the matricellular protein hevin in mouse brain is dependent on ADAMTS4.
    Authors: Weaver MS, Workman G, Cardo-Vila M, Arap W, Pasqualini R, Sage EH
    J. Biol. Chem., 2010;285(8):5868-77.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Bioassay
Primary Antibodies
Description Application Cat# Citations Images  

His Tag Antibody

WB, Simple Western, Flow, AP, CyTOF-ready MAB050 26  
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His Tag HRP-conjugated Antibody

WB, Simple Western MAB050H 3  
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His Tag APC-conjugated Antibody

ICFlow IC050A  
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His Tag Biotinylated Antibody

WB BAM050 4  
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His Tag PE-conjugated Antibody

ICFlow IC050P 2  
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His Tag Antibody

WB, Flow, AP MAB050R  
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His Tag Alexa Fluor® 488-conjugated Antibody

ICFlow IC050G  
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His Tag PerCP-conjugated Antibody

ICFlow IC050C  
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His Tag Alexa Fluor® 700-conjugated Antibody

ICFlow IC050N  
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Protein Purification Kits
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Histidine-tagged Protein Purification Resin

IP999 1
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Recombinant Proteins
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Recombinant Human Aggrecan G1-IGD-G2 Domains Protein, CF

Bind, EnzAct 1220-PG 1
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