Recombinant Human Nephrin Protein, CF

Catalog # Availability Size / Price Qty
9399-NN-050
Recombinant Human Nephrin Protein Bioactivity
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Recombinant Human Nephrin Protein, CF Summary

Product Specifications

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

Product Datasheets

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9399-NN

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

9399-NN

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

Scientific Data

Bioactivity Recombinant Human Nephrin Protein Bioactivity View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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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.
Entrez Gene IDs
4868 (Human); 54631 (Mouse); 64563 (Rat)
Alternate Names
CNF; Nephrin; nephrosis 1, congenital, Finnish type (nephrin); NPHNCNF; NPHS1; Renal glomerulus-specific cell adhesion receptor

Citation for Recombinant Human Nephrin Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Evaluation of Methodologies in Anti-nephrin Autoantibody Detection
    Authors: Liu, P;Liu, S;Dalal, V;Lane, J;Gessaroli, E;Forte, E;Gallon, L;Jin, J;
    bioRxiv : the preprint server for biology
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

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