Recombinant Human Nephrin Protein, CF

R&D Systems, part of Bio-Techne | Catalog # 9399-NN

R&D Systems, part of Bio-Techne
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9399-NN-050
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Key Product Details

Source

NS0

Accession #

O60500

Conjugate

Unconjugated

Applications

Bioactivity

View all Nephrin Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Nephrin protein
Gln23-Thr1029, with a C-terminal 6-His tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

No results obtained. Gln23 inferred from enzymatic pyroglutamate treatment revealing Leu24.

Predicted Molecular Mass

110 kDa

SDS-PAGE

119-155 kDa, reducing conditions

Activity

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.

Scientific Data Images for Recombinant Human Nephrin Protein, CF

Recombinant Human Nephrin Protein Bioactivity

Recombinant Human Nephrin Protein Bioactivity

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

Formulation, Preparation and Storage

9399-NN
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution
Reconstitute at 500 μg/mL in PBS.

Reconstitution Buffer Available:
Size / Price
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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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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 Recombinant Human Nephrin Protein, CF

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Product Specific Notices for Recombinant Human Nephrin Protein, CF

For research use only

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