Human ADAMTS4 Antibody Summary
Phe213-Cys685
Accession # O75173
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of ADAMTS4 by Western Blot MiR-23b-3p inhibits HS6ST2 expression by targeting HS6ST2 mRNA and effects specific gene expression in chondrocytes. A, b Dual luciferase reporter assay to validate target relationship between HS6ST2 and miR-23b-3p. SW1353 cells were transfected with mimic miR-23b-3p (a) or anti-miR-23b-3p (b) by control vector, pmirGLO-HS6ST2 wild-type 3′UTR vector or pmirGLO-HS6ST2 mutant 3′UTR vector, respectively, for 48 h. c, d Stem-loop RT-qPCR and western blotting results in SW1353 cells (c) or C28/I2 cells (d) transfected with 10 nM mimic miR-23b-3p (left panel) or 50 nM anti-miR-23b-3p sequence (right panel). e, f RT-qPCR (e) and western blotting (f) results of cartilage-specific gene expression in SW1353 cells transfected with mimic miR-23b-3p or negative control. g, h RT-qPCR (g) and western blotting (h) results of cartilage-specific gene expression in SW1353 cells transfected with anti-miR-23b-3p or negative control. RNA was harvested at 24 h, while protein was isolated at 48 h after transfection. U6 snRNA was used as internal controls in miRNA stem-loop RT-qPCR detection and GAPDH was used as internal controls in mRNA RT-qPCR and western blotting detection. Bars represent standard error of the mean (SEM) from three independent experiments. Mann–Whitney U test was used to identify statistical differences between two groups. *P value < 0.05 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29899528), licensed under a CC-BY license. Not internally tested by R&D Systems.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
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 (R&D Systems, Catalog # 973-TM).
- Totorella, M. D. et al. (1999) Science 284:1664.
- Porter, S. et al. (2005) Biochem. J. 386:15.
- Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
- Tortorella, M. D. et al. (2004) J. Biol. Chem. 279:17554.
- Struglics, A. et al. (2006) Osteoarthritis Cartilage. 14:101.
- Kashiwagi, M. et al. (2001) J. Biol. Chem. 276:12501.
- Pratta, M. A. et al. (2003) Arthritis Reum. 48:119.
- Glasson, S. S. et al. (2004) Arthritis Reum. 50:2547.
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