Nephrin Antibody [Unconjugated]

Nephrin was detected in perfusion fixed frozen sections of mouse kidney using Goat Anti-Mouse Nephrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3159) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Nephrin was detected in perfusion fixed frozen sections of mouse kidney using 10 µg/mL Goat Anti-Mouse Nephrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3159) overnight at 4 °C. Tissue was stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained (green). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

• Reactivity: Mu
• Applications: IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Nephrin Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Nephrin
  Gln37-Thr1049
  Accession # NP_062332
• Specificity: Detects mouse Nephrin in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant human Nephrin is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: NPHS1

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 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 [Unconjugated]

• 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. USA 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.

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.

Publications for Nephrin Antibody (AF3159)(11)

Publications using AF3159

• S Alarcón, W Garrido, C Cappelli, R Suárez, C Oyarzún, C Quezada, R San Martín Deficient Insulin-mediated Upregulation of the Equilibrative Nucleoside Transporter 2 Contributes to Chronically Increased Adenosine in Diabetic Glomerulopathy Sci Rep, 2017;7(1):9439. 2017 [PMID: 28842605] (IHC,, Rat)
• Y Kato, K Mori, M Kasahara, K Osaki, A Ishii, KP Mori, N Toda, S Ohno, T Kuwabara, T Tokudome, I Kishimoto, MA Saleem, T Matsusaka, K Nakao, M Mukoyama, M Yanagita, H Yokoi Natriuretic peptide receptor guanylyl cyclase-A pathway counteracts glomerular injury evoked by aldosterone through p38 mitogen-activated protein kinase inhibition Sci Rep, 2017;7(0):46624. 2017 [PMID: 28429785] (IHC,, Mouse)
• SJ Sung, Y Ge, C Dai, H Wang, SM Fu, R Sharma, YS Hahn, J Yu, TH Le, MD Okusa, WK Bolton, JR Lawler Dependence of Glomerulonephritis Induction on Novel Intraglomerular Alternatively Activated Bone Marrow-Derived Macrophages and Mac-1 and PD-L1 in Lupus-Prone NZM2328 Mice J. Immunol, 2017;0(0):. 2017 [PMID: 28219886] (Flow,, Mouse)
• Z Mohammed-A, C Lu, MK Marway, RE Carlisle, K Ask, D Lukic, JC Krepinsky, JG Dickhout Endoplasmic reticulum stress inhibition attenuates hypertensive chronic kidney disease through reduction in proteinuria Sci Rep, 2017;7(0):41572. 2017 [PMID: 28148966] (Western Blot,, Human)
• Alteration of histone H3K4 methylation in glomerular podocytes associated with proteinuria in patients with membranous nephropathy BMC Nephrol, 2016;17(1):179. 2016 [PMID: 27855638] (In vivo,, Mouse)
• Sun Y, Qu X, Howard V, Dai L, Jiang X, Ren Y, Fu P, Puelles V, Nikolic-Paterson D, Caruana G, Bertram J, Sleeman M, Li J Smad3 deficiency protects mice from obesity-induced podocyte injury that precedes insulin resistance. Kidney Int, 2015;88(2):286-98. 2015 [PMID: 25945408] (WB,, Mouse)
• Kim S, Lee S, Wang Z, Ding Y, Haque N, Zhang J, Zhou J, Choi M TGF-beta-activated kinase 1 is crucial in podocyte differentiation and glomerular capillary formation. J Am Soc Nephrol, 2014;25(9):1966-78. 2014 [PMID: 24652804] (IHC,, Human)
• Lan, Xiqian, Rai, Partab, Chandel, Nirupama, Cheng, Kang, Lederman, Rivka, Saleem, Moin A, Mathieson, Peter W, Husain, Mohammad, Crosson, John T, Gupta, Kalpna, Malhotra, Ashwani, Singhal, Pravin C Morphine induces albuminuria by compromising podocyte integrity. PLoS ONE, 2013;8(3):e55748. 2013 [PMID: 23555556] (IHC Paraffin-embedded,, Mouse)
• Dane M, van den Berg B, Avramut M, Faas F, van der Vlag J, Rops A, Ravelli R, Koster B, van Zonneveld A, Vink H, Rabelink T Glomerular endothelial surface layer acts as a barrier against albumin filtration. Am J Pathol, 2013;182(5):1532-40. 2013 [PMID: 23518410] (IHC Frozen,, Mouse)
• Divergent roles of Smad3 and PI3-kinase in murine adriamycin nephropathy indicate distinct mechanisms of proteinuria and fibrogenesis. Kidney Int., 2012;82(5):525-36. 2012 [PMID: 22534961] (IHC Paraffin-embedded,, Mouse)
• Pierchala BA, Munoz MR, Tsui CC Proteomic analysis of the slit diaphragm complex: CLIC5 is a protein critical for podocyte morphology and function. Kidney Int., 2010;78(9):868-82. 2010 [PMID: 20664558] (IHC,, Mouse)

Bioinformatics

• Gene Symbol: NPHS1
• Entrez:
  • 4868(Human)
  • 54631(Mouse)
  • 64563(Rat)
• Uniprot:
  • NP_062332()