Nephrin Antibody (2795E) [Unconjugated]

Nephrin was detected in immersion fixed paraffin-embedded sections of human kidney using Rabbit Anti-Human Nephrin Monoclonal Antibody (Catalog # MAB42693) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to glomeruli. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu
• Applications: IHC
• Clone: 2795E
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

Nephrin Antibody (2795E) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Mouse myeloma cell line NS0-derived human Nephrin protein
  Gln23-Thr1029
  Accession # O60500
• Specificity: Detects human Nephrin in direct ELISAs.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-25 ug/mL

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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

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

Alternate Names for Nephrin Antibody (2795E) [Unconjugated]

• 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • O60500()