Recombinant Human Nephrin Protein, CF

  
  • Purity
    >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
  • 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 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
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.
Data Images
When Recombinant Human Podocin/NPHS2 (Catalog # 9287-PO) is coatedonto a microplate at 2 μg/mL, Recombinant Human Nephrin (Catalog # 9399-NN) bindswith an ED50 of 0.3-1.5 µg/mL.
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
Related Research Areas
Primary Antibodies
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His Tag Antibody

WB, Flow, AP MAB050R  
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His Tag Antibody

WB, Simple Western, Flow, AP, CyTOF-ready MAB050 26  
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His Tag HRP-conjugated Antibody

WB, Simple Western MAB050H 3  
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His Tag APC-conjugated Antibody

ICFlow IC050A  
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His Tag PE-conjugated Antibody

ICFlow IC050P 2  
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His Tag Biotinylated Antibody

WB BAM050 3  
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His Tag Alexa Fluor® 488-conjugated Antibody

ICFlow IC050G  
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His Tag PerCP-conjugated Antibody

ICFlow IC050C  
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His Tag Alexa Fluor® 700-conjugated Antibody

ICFlow IC050N  
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His Tag Alexa Fluor® 700-conjugated Antibody

ICFlow IC0501N
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His Tag Alexa Fluor® 405-conjugated Antibody

ICFlow IC0501V
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His Tag Alexa Fluor® 750-conjugated Antibody

ICFlow IC0501S
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His Tag Alexa Fluor® 350-conjugated Antibody

ICFlow IC050U
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His Tag Alexa Fluor® 594-conjugated Antibody

ICFlow IC050T
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His Tag Alexa Fluor® 488-conjugated Antibody

ICFlow IC0501G
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His Tag Alexa Fluor® 405-conjugated Antibody

ICFlow IC050V
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His Tag Alexa Fluor® 647-conjugated Antibody

ICFlow IC050R
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His Tag Alexa Fluor® 647-conjugated Antibody

ICFlow IC0501R
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His Tag Alexa Fluor® 750-conjugated Antibody

ICFlow IC050S
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His Tag Alexa Fluor® 594-conjugated Antibody

ICFlow IC0501T
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His Tag Alexa Fluor® 350-conjugated Antibody

ICFlow IC0501U
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Protein Purification Kits
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Histidine-tagged Protein Purification Resin

IP999 1
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