Recombinant Mouse Nephrin Protein, CF

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

Recombinant Mouse Nephrin Protein, CF Summary

• Details of Functionality: Measured by the ability of the immobilized protein to support the adhesion of MS‑1 mouse pancreatic islet endothelial cells. When 5 x 10⁴ cells/well are added to rmNephrin coated plates (10 µg/mL, 100 µL/well), >60% will adhere after 90 minutes at 37° C.
• Source: Mouse myeloma cell line, NS0-derived mouse Nephrin protein
  Gln37-Thr1049, with a C-terminal 6-His tag
• Accession #: NP_062332
• N-terminal Sequence: No results obtained: Gln37 predicted
• Protein/Peptide Type: Recombinant Proteins
• Gene: Nphs1
• Purity: >85%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 110 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: 130-145 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 PBS.
• Purity: >85%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• 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 Nephrin Protein, CF

• CNF
• Nephrin
• nephrosis 1, congenital, Finnish type (nephrin)
• NPHNCNF
• NPHS1
• Renal glomerulus-specific cell adhesion receptor

Background

Nephrin is a 185 kDa type I transmembrane glycoprotein that belongs to the immunoglobulin superfamily (1). Mature mouse Nephrin consists of a 1042 amino acid (aa) extracellular domain (ECD) with eight Ig-like C2-set domains and one fibronectin type III domain, a 22 aa transmembrane segment, and a 156 aa cytoplasmic tail (2, 3). Within the ECD, mouse Nephrin shares 84% and 95% aa sequence identity with human and rat Nephrin, respectively. Usage of the alternate exon 1B results in a distinct N-terminal sequence that lacks a clearly defined signal peptide cleavage site (4). Nephrin is expressed primarily on podocytes in the renal glomerulus and to a lesser extent in the brain and pancreas (3, 5). The 1B isoform is not expressed in the kidney (4). Nephrin localizes to intercellular junctions between podocyte foot processes where it functions as a homophilic adhesion molecule (2, 6). Nephrin is required for formation and maintenance of the slit diaphragm between these processes (7). It associates with Neph1, podicin, P-cadherin, and multiple scaffolding proteins which couple it to the actin cytoskeleton (8 - 12). Nephrin expression is required for the anti-apoptotic effect of VEGF on podocytes as well as for the ability of podocytes to upregulate Glut1 and Glut4 glucose transporters in response to insulin (13, 14). Nephrin downregulation contributes to diabetic nephropathy, and Nephrin mutations underlie the lethal congenital nephritic syndrome NPHS1 (5, 15).

1. Kawachi, H. et al. (2006) Nephrology 11:274.
2. Holzman, L.B. et al. (1999) Kidney Int. 56:1481.
3. Putaala, H. et al. (2000) J. Am. Soc. Nephrol. 11:991.
4. Beltcheva, O. et al. (2003) J. Am. Soc. Nephrol. 14:352.
5. Putaala, H. et al. (2001) Hum. Mol. Genet. 10:1.
6. Khoshnoodi, J. et al. (2003) Am. J. Pathol. 163:2337.
7. Ruotsalainen, V. et al. (2000) Am. J. Pathol. 157:1905.
8. Barletta, G.M. et al. (2003) J. Biol. Chem. 278:19266.
9. Huber, T.B. et al. (2001) J. Biol. Chem. 276:41543.
10. Lehtonen, S. et al. (2004) Am. J. Pathol. 165:923.
11. Lehtonen, S. et al. (2005) Proc. Natl. Acad. Sci. 102:9814.
12. Verma, R. et al. (2006) J. Clin. Invest. 116:1346.
13. Foster, R.R. et al. (2005) Am. J. Physiol. Renal Physiol. 288:F48.
14. Coward, R.J. et al. (2007) Diabetes 56:1127.
15. Cooper, M.E. et al. (2002) Semin. Nephrol. 22:393.

Bioinformatics

• Gene Symbol: Nphs1
• Uniprot:
  • NP_062332()