Anti-Neph1 Antibody, cytoplasmic domain

Kin of IRRE-like protein 1 (UniProt Q80W68; also known as Nephrin-like protein 1, Kin of IRRE like 1, Kin of irregular chiasm-like protein 1) is encoded by the Kirrel (also known as 6720469N11Rik, Kirrel1, Neph1) gene (Gene ID 170643) in murine species. Podocytes are specialized epithelial cells that are critical components of the glomerular filtration barrier. Glomerular injury, such as that caused by renal ischemia, is often characterized by the effacement of podocytes, loss of slit diaphragms, and proteinuria. The podocyte proteins Neph1 and nephrin are critical for maintaining the structural integrity of slit diaphragm and therefore the filtration function of the glomerulus by organizing a signaling complex at the podocyte cell membrane. Cytoplasmic domain of Neph1 plays a key role in actin cytoskeleton remodeling events that are directly associated with Fyn-mediated Neph1 pTyr637/638 phosphorylation that leads to the recruitment of adapter proteins and signaling molecules, such as Grb2, Csk tyrosine kinase, and ZO-1. Phosphorylation-induced association between Neph1 and ZO-1 plays a key role in defining the tight junction formation in podocytes.

Expected to react with murine spliced isoforms 1&2, as well as all three human spliced isoforms.

Epitope: cytoplasmic domain

His-tagged recombinant protein corresponding to the cytoplasmic domain of mouse Neph1.

Detect Neph1 using this rabbit Polyclonal Anti-Neph1 Antibody, cytoplasmic domain, Cat. No. ABS1511, validated for use in Western Blotting, Immunoprecipitation, Immunofluorescence, Electron Microscopy and Immunocytochemistry.

Research Category
Signaling

Research Sub Category
Signaling Neuroscience

Western Blotting Analysis: A representative lot detected endogenous rat glomeruli Neph1 and exogenously expressed mouse Neph1 in COS-7 cells (Barletta, G.M., et al. (2003). J Biol Chem. 278(21):19266-19271).
Western Blotting Analysis: A representative lot detected ischemia-induced Neph1 membrane-to-cytosol translocation in human podocytes (Wagner, M.C., et al. (2008). J Biol Chem. 283(51):35579-35589).
Western Blotting Analysis: A representative lot detected Neph1 in mouse glomeruli & cultured human podocytes (Arif, E., et al. (2011). Mol Cell Biol. 31(10):2134-2150).
Immunoprecipitation Analysis: A representative lot immunoprecipitated Neph1 from rat glomerular and human podocyte cell lysates (Arif, E., et al. (2011). Mol Cell Biol. 31(10):2134-2150).
Immunofluorescence Analysis: A representative lot detected Neph1 using both paraffin-embedded and frozen rat kidney sections (Arif, E., et al. (2011). Mol Cell Biol. 31(10):2134-2150; Barletta, G.M., et al. (2003). J Biol Chem. 278(21):19266-19271).
Electron Microscopy Analysis: A representative lot detected Neph1 in frozen rat kidney sections (Barletta, G.M., et al. (2003). J Biol Chem. 278(21):19266-19271).
Immunocytochemistry Analysis: A representative lot detected Neph1 in cultured human podocytes (Arif, E., et al. (2014). J Biol Chem. 289(14):9502-9518; Arif, E., et al. (2011). Mol Cell Biol. 31(10):2134-2150; Wagner, M.C., et al. (2008). J Biol Chem. 283(51):35579-35589).

Evaluated by Western Blotting in rat kidney tissue lysate.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Neph1 in rat kidney tissue lysate.

~110 kDa observed. Target band size appears larger than the calculated molecular weight (87.2 kDa/murine isoform 1, 70.0 kDa/murine isoform 2, 72.4-85.0 kDa/human isoform 1-3) due to posttranslational modifications (glycosylation & phosphorylation). An uncharacterized band ~25 kDa can also be seen in some samples.

Affinity purified

Purified rabbit polyclonal antibody in buffer containing PBS/glycine/5% BSA, .02% Sodium Azide and 30% glycerol.

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Concentration: Please refer to lot specific datasheet.

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