Recombinant Human Nephrin Protein, CF

When Recombinant Human Podocin/NPHS2 (Catalog # 9287-PO) is coatedonto a microplate at 2 μg/mL, Recombinant Human Nephrin (Catalog # 9399-NN) bindswith an ED50 of 0.3-1.5 µg/mL.

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

Recombinant Human Nephrin Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human Podocin/NPHS2 (Catalog # 9287-PO) is immobilized at 2 μg/mL, 100 μL/well, the concentration of Recombinant Human Nephrin that produces 50% of the optimal binding response is 0.3-1.5 μg/mL.
• Source: Mouse myeloma cell line, NS0-derived human Nephrin protein
  Gln23-Thr1029, with a C-terminal 6-His tag
• Accession #: O60500
• N-terminal Sequence: No results obtained. Gln23 inferred from enzymatic pyroglutamate treatment revealing Leu24.
• Protein/Peptide Type: Recombinant Proteins
• 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: 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: 119-155 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 with Trehalose.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant Human Nephrin Protein, CF

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

Background

Nephrin is a 180 kDa type I transmembrane glycoprotein that belongs to the immunoglobulin superfamily (1). Mature human Nephrin consists of a 1033 amino acid (aa) extracellular domain (ECD) with eight Ig-like C2-set domains and one fibronectin type III domain, a 21 aa transmembrane segment, and a 165 aa cytoplasmic tail (2, 16). Within the ECD, human Nephrin shares 83% aa sequence identity with both mouse and rat Nephrin (3). 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 up-regulate Glut1 and Glut4 glucose transporters in response to insulin (13, 14). Nephrin down-regulation contributes to diabetic nephropathy, and nephrin mutations underlie the lethal congenital nephritic syndrome NPHS1 (5, 15).

1. Ruotsalainen, V. et al. 1999, Proc. Natl. Acad. Sci. USA 96: 7962.
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.
16. Kestila¨, M. et al. 1998, Mol. Cell. 1:575.

Publications for Nephrin (9399-NN)(1)

Publication using 9399-NN

• Liu, P;Liu, S;Dalal, V;Lane, J;Gessaroli, E;Forte, E;Gallon, L;Jin, J; Evaluation of Methodologies in Anti-nephrin Autoantibody Detection bioRxiv : the preprint server for biology 2024-07-25 [PMID: 39211159] (Bioassay, Human)

Bioinformatics

• Uniprot:
  • O60500()