Recombinant Human Nephrin Protein, CF Summary
Product Specifications
Gln23-Thr1029, with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
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Scientific Data
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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
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).
- Ruotsalainen, V. et al. 1999, Proc. Natl. Acad. Sci. USA 96: 7962.
- Holzman, L.B. et al. 1999, Kidney Int. 56:1481.
- Putaala, H. et al. 2000, J. Am. Soc. Nephrol. 11:991.
- Beltcheva, O. et al. 2003, J. Am. Soc. Nephrol. 14:352.
- Putaala, H. et al. 2001, Hum. Mol. Genet. 10:1.
- Khoshnoodi, J. et al. 2003, Am. J. Pathol. 163:2337.
- Ruotsalainen, V. et al. 2000, Am. J. Pathol. 157:1905.
- Barletta, G.M. et al. 2003, J. Biol. Chem. 278:19266.
- Huber, T.B. et al. 2001, J. Biol. Chem. 276:41543.
- Lehtonen, S. et al. 2004, Am. J. Pathol. 165:923.
- Lehtonen, S. et al. 2005, Proc. Natl. Acad. Sci. 102:9814.
- Verma, R. et al. 2006, J. Clin. Invest. 116:1346.
- Foster, R.R. et al. 2005, Am. J. Physiol. Renal Physiol. 288:F48.
- Coward, R.J. et al. 2007, Diabetes 56:1127.
- Cooper, M.E. et al. 2002, Semin. Nephrol. 22:393.
- Kestila¨, M. et al. 1998, Mol. Cell. 1:575.
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