Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2795E
View all Nephrin Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived human Nephrin protein
Gln23-Thr1029
Accession # O60500

Specificity

Detects human Nephrin in direct ELISAs.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human Nephrin Antibody

Nephrin in Human Kidney.

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.

Applications for Human Nephrin Antibody

Application
Recommended Usage

Immunohistochemistry

3-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human kidney

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Nephrin

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).

References

  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.

Alternate Names

CNF, NPHS1

Entrez Gene IDs

4868 (Human); 54631 (Mouse); 64563 (Rat)

Gene Symbol

NPHS1

UniProt

O60500

Additional Nephrin Products

Product Documents for Human Nephrin Antibody

Certificate of Analysis

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Product Specific Notices for Human Nephrin Antibody

For research use only

Related Research Areas

Immunoglobulin Superfamily CAMs

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Protocols

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