Recombinant Human Thrombospondin-4 Protein, CF

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Thrombospondin-4 Protein, CF Summary

• Details of Functionality: Measured by the ability of the immobilized protein to support the adhesion of SVEC4‑10 mouse vascular endothelial cells. Recombinant Human Thrombospondin‑4 (rhThrombospondin-4) is coated to 96 well plates at 10 µg/mL (100 µL/well) overnight then reduced with 20 mM DTT for 30 minutes. When 2 x 10⁴ cells/well are added to rhThrombospondin‑4 coated plates, ≥50% will adhere after one hour at 37 °C.
  Optimal dilutions should be determined by each laboratory for each application.
• Source: Chinese Hamster Ovary cell line, CHO-derived human Thrombospondin-4 protein
  Ala22-Asn961, with a C-terminal 10-His tag
• Accession #: P35443
• N-terminal Sequence: Ala22
• Protein/Peptide Type: Recombinant Proteins
• Gene: THBS4
• 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

• Dilutions:
  • Bioactivity
• Theoretical MW: 105 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: 145 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile PBS.

Notes

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

Alternate Names for Recombinant Human Thrombospondin-4 Protein, CF

• THBS4
• thrombospondin 4
• Thrombospondin4
• Thrombospondin-4
• TSP4
• TSP-4

Background

Thrombospondin-4 (TSP-4) is an approximately 140 kDa matricellular protein that is secreted as a disulfide-linked pentamer. Within the Thrombospondin family, TSP-3 and TSP-5/COMP are also pentameric, while TSP-1 and TSP-2 are trimeric. TSP-4 regulates cell-cell and cell matrix interactions and plays a role in cardiovascular physiology and neuronal development (1, 2). Mature human TSP-4 consists of an N-terminal heparin-binding domain, a coiled coil motif, four EGF-like repeats, seven TSP type-3 repeats (one with an RGD motif), and a TSP C-terminal domain (2). Human TSP-4 shares 93% amino acid sequence identity with mouse and rat TSP-4. TSP-4 binds a variety of matrix proteins including Collagens I, II, III, V, Laminin-1, Fibronectin, and Matrilin-2 (3). Interactions of TSP-4 with non-collagenous proteins are independent of divalent cations, whereas interactions with collagenous proteins are enhanced in the presence of zinc (3). TSP-4 binds to cell surface Integrins containing the alpha M, beta 2, or beta 3 chains (4, 5). It is expressed in skeletal muscle and tendon as well as by vascular smooth muscle and endothelial cells (6‑8). It is up‑regulated in cardiomyocytes during pressure overload and is required for mediating the responsive increase in cardiac contractility (9). In humans, a polymorphism of TSP-4 (A387P) is associated with myocardial infarction (10). TSP-4 contributes to the development of inflammation and atherosclerosis by promoting macrophage and neutrophil adhesion to the vasculature (4, 5). In the nervous system, TSP-4 is expressed by astrocytes and neurons and is enriched at neuromuscular junctions and synapse-rich layers of the brain and retina (8, 11, 12). It promotes neuronal adhesion, neurite outgrowth, and excitatory synaptogenesis (8, 12, 13). TSP-4 is
up‑regulated in the spinal cord following peripheral nerve injury where it contributes to presynaptic hypersensitivity and hyperalgesia (11). It is also up‑regulated in muscle following denervation (8). TSP-4 is additionally secreted by tumor-associated fibroblasts in gastric adenocarcinoma, particularly in regions of tumor cell invasion (14).

1. Murphy-Ullrich, J.E. and R.V. Iozzo (2012) Matrix Biol. 31:152.
2. Lawler, J. et al. (1995) J. Biol. Chem. 270:2809.
3. Narouz-Ott, L. et al. (2000) J. Biol. Chem. 275:37110.
4. Pluskota, E. et al. (2005) Blood 106:3970.
5. Frolova, E.G. et al. (2010) Circ. Res. 107:1313.
6. Sodersten, F. et al. (2007) Connect. Tiss. Res. 48:254.
7. Stenina, O.I. et al. (2003) Circulation 108:1514.
8. Arber, S. and P. Caroni (1995) J. Cell Biol. 131:1083.
9. Cingolani, O.H. et al. (2011) Circ. Res. 109:1410.
10. Stenina, O.I. et al. (2007) Arterioscler. Thromb. Vasc. Biol. 27:1886.
11. Kim, D.-S. et al. (2012) J. Neurosci. 32:8977.
12. Dunkle, E.T. et al. (2007) Exp. Eye Res. 84:707.
13. Eroglu, C. et al. (2009) Cell 139:380.
14. Forster, S. et al. (2011) Mod. Pathol. 24:1390.

Publications for Thrombospondin-4 (2390-TH)(1)

Publication using 2390-TH

• Jeschke A, Bonitz M, Simon M, Peters S, Baum W, Schett G, Ruether W, Niemeier A, Schinke T, Amling M Deficiency of Thrombospondin-4 in Mice Does Not Affect Skeletal Growth or Bone Mass Acquisition, but Causes a Transient Reduction of Articular Cartilage Thickness. PLoS ONE, 2015-12-02;10(12):e0144272. 2015-12-02 [PMID: 26629997] (Bioassay, Porcine)

Bioinformatics

• Gene Symbol: THBS4
• Uniprot:
  • P35443()