Recombinant Mouse Nephronectin Protein, CF

Recombinant Mouse Nephronectin (Catalog # 4298-NP) supports the adhesion of SVEC4‑10mouse vascular endothelial cells. When 5 x 104cells/well are added to Recombinant Mouse Nephronectin coated plates (10 µg/mL, 100 µL/well), approximately 50-80% will adhere after 1hour at 37 °C.

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

Recombinant Mouse Nephronectin 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. When 5 x 10⁴ cells/well are added to rmNephronectin coated plates (10 µg/mL,
  100 µL/well), approximately 50-80% will adhere after 1 hour at 37 °C.
• Source: Mouse myeloma cell line, NS0-derived mouse Nephronectin protein
  Asp20-Cys561, with a C-terminal 6-His tag
• Accession #: AAH68308
• N-terminal Sequence: Asp20
• Protein/Peptide Type: Recombinant Proteins
• Gene: Npnt
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 60.3 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: 80-95 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 PBS.
• 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 Mouse Nephronectin Protein, CF

• EGFL6L
• Nctn
• Nephronectin
• NPNT
• POEM

Background

Nephronectin, also named POEM (preosteoblast EGF repeat protein with MAM domain) is a 70 - 90 kDa extracellular matrix protein that is a ligand for integrin alpha ₈ beta ₁ (1, 2). The 561 amino acid (aa) mouse Nephronectin (isoform b) contains a 19 aa signal sequence, a matrilin-type vWA domain, two calcium-binding EGF-like domains, a potential N-linked glycosylation site, a pro/ser/thr-rich mucin-like region, an RGD integrin binding motif, and a MAM domain. These features are often present in matrix proteins that, like Nephronectin, mediate cell adhesion and spreading (1, 2). Isoform a includes a 17 aa insertion between aa 58 and 59 that is missing in isoform b. Mouse Nephronectin shows 88%, 92%, 85% and 77% aa identity with human, rat, bovine and opossum Nephronectin, respectively. It is most highly expressed in developing endocrine organs such as parathyroid, thyroid, hypophysis and pineal organ, and around developing bone, teeth, and muscle (1, 2). Pre-osteoblast cells produce Nephronectin, but downregulate it as they differentiate (1). It is also expressed in the Wolffian duct and ureteric bud basement membranes in the developing kidney, where it is colocalizes, and forms an in-vivo complex with integrin alpha ₈ beta ₁ (2). Since deletion of integrin alpha ₈ beta ₁ results in renal agenesis, Nephronectin has been proposed to be a critical integrin alpha ₈ beta ₁ ligand during Nephrogenesis (2, 3).

1. Morimura, N. et al. (2001) J. Biol. Chem. 276:42172.
2. Brandenberger, R. et al. (2001) J. Cell Biol. 154:447.
3. Miner, J. H. (2001) J. Cell Biol. 154:257.

Publications for Nephronectin (4298-NP)(8)

Publications using 4298-NP

• J Basta, L Robbins, L Stout, MJ Prinsen, DW Griggs, M Rauchman Pharmacologic inhibition of RGD-binding integrins ameliorates fibrosis and improves function following kidney injury Physiol Rep, 2020-04-01;8(7):e14329. 2020-04-01 [PMID: 32281744] (Bioassay, Human)
• S Kon, M Honda, K Ishikawa, M Maeda, T Segawa Antibodies against nephronectin ameliorate anti-type II collagen-induced arthritis in mice FEBS Open Bio, 2019-11-24;0(0):. 2019-11-24 [PMID: 31705832] (ELISA (Standard), Mouse)
• CF Hung, CL Wilson, YH Chow, LM Schnapp Role of integrin alpha8 in murine model of lung fibrosis PLoS ONE, 2018-05-29;13(5):e0197937. 2018-05-29 [PMID: 29813125] (Bioassay, Mouse)
• TS Steigedal, J Toraskar, RP Redvers, M Valla, SN Magnussen, AM Bofin, S Opdahl, S Lundgren, BL Eckhardt, JM Lamar, J Doherty, RO Hynes, RL Anderson, G Svineng Nephronectin is Correlated with Poor Prognosis in Breast Cancer and Promotes Metastasis via its Integrin-Binding Motifs Neoplasia, 2018-03-11;20(4):387-400. 2018-03-11 [PMID: 29539586] (Bioassay, Mouse)
• J Tang, T Saito The Role of Nephronectin on Proliferation and Differentiation in Human Dental Pulp Stem Cells Stem Cells Int, 2017-11-19;2017(0):2546261. 2017-11-19 [PMID: 29358954] (Bioassay, Human)
• AE Teo, S Garg, TI Johnson, W Zhao, J Zhou, CE Gomez-Sanc, M Gurnell, MJ Brown Physiological and Pathological Roles in Human Adrenal of the Glomeruli-Defining Matrix Protein NPNT (Nephronectin) Hypertension, 2017-04-17;0(0):. 2017-04-17 [PMID: 28416583] (Bioassay, Human)
• C Arai, K Yoshizaki, K Miyazaki, K Saito, A Yamada, X Han, K Funada, E Fukumoto, N Haruyama, T Iwamoto, I Takahashi, S Fukumoto Nephronectin plays critical roles in Sox2 expression and proliferation in dental epithelial stem cells via EGF-like repeat domains Sci Rep, 2017-03-27;7(0):45181. 2017-03-27 [PMID: 28345658] (Bioassay, Mouse)
• Zhang H, Tasnim F, Ying JY, Zink D The impact of extracellular matrix coatings on the performance of human renal cells applied in bioartificial kidneys. Biomaterials, 2009-02-12;30(15):2899-911. 2009-02-12 [PMID: 19217158] (Bioassay, Human)

Bioinformatics

• Gene Symbol: Npnt
• Uniprot:
  • AAH68308()