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Recombinant Human ADAMTS5 Protein, CF

Catalog #: 2198-AD
11 Citations 1 Review Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
2198-AD-020
R&D Systems Recombinant Proteins and Enzymes
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Citations (11)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human ADAMTS5 Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
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 >10 pmol/min/μg, as measured under the described conditions. 
Source
Mouse myeloma cell line, NS0-derived human ADAMTS5 protein
Ser262-Pro622, with a C-terminal 10-His tag
Accession #
Q9UNA0
N-terminal Sequence
Analysis
Ser262
Structure / Form
Recombinant Human ADAMTS5 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
Predicted Molecular Mass
41 kDa
SDS-PAGE
51 kDa, reducing conditions

Product Datasheets

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2198-AD

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

2198-AD

Formulation Lyophilized from a 0.2 μm filtered solution in Sodium Acetate, CaCl2 and NaCl.
Reconstitution Reconstitute at 200 μg/mL in sterile, deionized water.
Shipping The product is shipped at ambient temperature. 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, -70 °C under sterile conditions after reconstitution.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 100 mM NaCl, 5 mM CaCl2, 0.05% Brij-35, pH 7.5
  • Recombinant Human ADAMTS5 (rhADAMTS5) (Catalog # 2198-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 rhADAMTS5 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 rhADAMTS5 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:
  • rhADAMTS5: 0.5 µg
  • Substrate: 25 µM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: ADAMTS5

ADAMTS5 (a disintegrin and metalloproteinase with thrombospondin motifs 5), also known as aggrecanase-2 and ADAMTS11, is a member of the family of secreted zinc proteases with a multi-domain structure (1, 2). The protein precursors consist of signal peptide and following domains: pro, catalytic, disintegrin-like, TS type 1 motif, cysteine-rich, spacer and a variable number of TS type 1 motifs. ADAMTS5 is an active protease effectively cleaving alpha 2-macroglobulin (3), aggrecan (4), and brevican (5), and is inhibited by TIMP-3 with inhibition constants in the subnanomolar range (6). Based on the murine model studies (7, 8), this protease may be a key enzyme in the degradation of cartilage leading to osteoarthritis and rheumatoid arthritis. The purified rhADAMTS5 starts at the N-terminus of the catalytic domain and ends at the C-terminus of the TSP-1 domain. The amino acid sequence of rhADAMTS5 is 98%, 97%, and 96% identical to that of canine, bovine, and mouse/rat. The aggrecanase activity can be inhibited by 5 mM 1,10-phenanthroline and recombinant human TIMP-3 (Catalog # 973-TM).

References
  1. Porter, S. et al. (2005) Biochem. J. 386:15.
  2. Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
  3. Tortorella, M.D. et al. (2004) J. Biol. Chem. 279:17554.
  4. Vankemmelbeke, M.N. et al. (2001) Eur. J. Biochem. 268:1259.
  5. Nakada, M. et al. (2005) Acta Neurophathol (Berl). 110:239.
  6. Kashiwagi, M. et al. (2001) J. Biol. Chem. 276:12501.
  7. Glasson, S.S. et al. (2005) Nature, 434: 644.
  8. Stanton, H. et al. (2005) Nature, 434:648.
Long Name
A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 5
Entrez Gene IDs
11096 (Human); 23794 (Mouse); 304135 (Rat)
Alternate Names
A disintegrin and metalloproteinase with thrombospondin motifs 11; a disintegrin-like and metalloprotease (reprolysin type) with thrombospondintype 1 motif, 5 (aggrecanase-2); ADAM metallopeptidase with thrombospondin type 1 motif, 5; ADAM-TS 11; ADAM-TS 5; ADAMTS11; ADAMTS-11; ADAMTS5; ADAM-TS5; ADAMTS-5; ADMP2; ADMP-2; ADMP-2A disintegrin and metalloproteinase with thrombospondin motifs 5; Aggrecanase 2; aggrecanase-2; disintegrin-like and metalloprotease with thrombospondin type 1 motif, 510ADAMTS11FLJ36738; EC 3.4.24; EC 3.4.24.-; EC 3.4.24.14; EC 3.4.24.82

Citations for Recombinant Human ADAMTS5 Protein, CF

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.

11 Citations: Showing 1 - 10
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  1. A plastic aggrecan barrier modulated by peripheral energy state gates metabolic signal access to arcuate neurons
    Authors: Kuczynski-Noyau, L;Karmann, S;Alberton, P;Martinez-Corral, I;Nampoothiri, S;Sauvé, F;Lhomme, T;Quarta, C;Apte, SS;Bouret, S;Aszodi, A;Rasika, S;Ciofi, P;Dam, J;Prévot, V;Mattot, V;
    Nature communications
    Species: Mouse
    Sample Types: In Vivo
    Applications: In vivo assay
  2. Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis
    Authors: DC Kirman, B Renganatha, WK Chui, MW Chen, NA Kaya, R Ge
    Cell Death & Disease, 2022-02-23;13(2):172.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. The Metalloproteinase ADAMTS5 Is Expressed by Interstitial Inflammatory Cells in IgA Nephropathy and Is Proteolytically Active on the Kidney Matrix
    Authors: S Taylor, M Whitfield, J Barratt, A Didangelos
    J. Immunol., 2020-09-11;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Bioassay
  4. Neurocan is a New Substrate for the ADAMTS12 Metalloprotease: Potential Implications in Neuropathies
    Authors: T Fontanil, Y Mohamedi, A Moncada-Pa, T Cobo, JA Vega, JL Cobo, O García-Suá, J Cobo, ÁJ Obaya, S Cal
    Cell. Physiol. Biochem., 2019-01-01;52(5):1003-1016.
    Species: Mouse
    Sample Types: Recombinant Protein
    Applications: Bioassay
  5. Upper zone of growth plate and cartilage matrix associated protein protects cartilage during inflammatory arthritis
    Authors: F Seuffert, D Weidner, W Baum, G Schett, M Stock
    Arthritis Res. Ther., 2018-05-02;20(1):88.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  6. ADAMTS5 Deficiency in Calcified Aortic Valves Is Associated With Elevated Pro-Osteogenic Activity in Valvular Interstitial Cells
    Authors: F Li, R Song, L Ao, TB Reece, JC Cleveland, N Dong, DA Fullerton, X Meng
    Arterioscler. Thromb. Vasc. Biol., 2017-05-25;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  7. 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, 2017-02-21;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  8. 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, 2016-01-25;11(1):10.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Bioassay
  9. The effect of protease inhibitors on the induction of osteoarthritis-related biomarkers in bovine full-depth cartilage explants.
    Authors: He Y, Zheng Q, Jiang M, Sun S, Christiansen T, Kassem M, Karsdal M, Bay-Jensen A
    PLoS ONE, 2015-04-24;10(0):.
    Applications: Western Blot
  10. ADAMTS5 functions as an anti-angiogenic and anti-tumorigenic protein independent of its proteoglycanase activity.
    Authors: Kumar S, Sharghi-Namini S, Rao N, Ge R
    Am J Pathol, 2012-07-13;181(3):1056-68.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  11. 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, 2012-06-11;287(32):26647-56.
    Applications: Enzyme Assay

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Recombinant Human ADAMTS5 Protein, CF
By Anonymous on 01/05/2018
Application: Characterization prior to use.
Other Recombinant Human ADAMTS5 Protein, CF 2198-AD