Recombinant Mouse Nephrin Protein, CF

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3159-NN-050
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Mouse Nephrin Protein, CF Summary

Product Specifications

Purity
>85%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by the ability of the immobilized protein to support the adhesion of MS‑1 mouse pancreatic islet endothelial cells. When 5 x 104 cells/well are added to rmNephrin coated plates (10 µg/mL, 100 µL/well), >60% will adhere after 90 minutes at 37° C.
Source
Mouse myeloma cell line, NS0-derived mouse Nephrin protein
Gln37-Thr1049, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
No results obtained: Gln37 predicted
Predicted Molecular Mass
110 kDa
SDS-PAGE
130-145 kDa, reducing conditions

Product Datasheets

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3159-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.

3159-NN

Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in sterile 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.
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Background: Nephrin

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).

References
  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. 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.
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

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