Anti-phospho-Neph1 (Tyr637/638) Antibody

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. The 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.

Mouse Neph1 Tyr637/638 site is equivalent to human Neph1 Tyr605/606. Equivalent target site is also present in human spliced isoforms 2&3 (Tyr621/622 & Tyr502/503, respectively), but not in murine spliced isoform 2.

Epitope: Cytoplasmic domain

Linear peptide corresponding to the cytoplasmic domain of mouse phospho-Neph1 (Tyr637/638).

Detect Neph1 using this rabbit Polyclonal Anti-phospho-Neph1 Antibody (Tyr637/638), Cat. No. ABS1509, validated for use in Western Blotting.

Research Category
Signaling

Research Sub Category
Signaling Neuroscience

Western Blotting Analysis: A representative lot detected Neph1 phosphorylation in kinase assay using Fyn and COS-7 lysate containing exogenously expressed Neph1 (Data courtesy of Deepak Nihalini, University of Pennsylvania)
Western Blotting Analysis: A representative lot detected Neph1 phosphorylation in lysates from puromycin aminonucleoside-/PAN-treated human pocodytes (Arif, E., et al. (2014). J Biol Chem. 289(14):9502-9518).

Evaluated by Western Blotting in pervanadate treated human podocyte cell lysate.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Neph1 phosphorylation at the target tyrosine residues in 7.5 µL of pervanadate treated human podocyte cell lysate.

~120 kDa observed. Target band size appears larger than the calculated molecular weight (87.2 kDa/murine isoform 1, 72.4-85.0 kDa/human isoform 1-3) due to posttranslational modifications (glycosylation & phosphorylation). Uncharacterized band(s) may appear in some lysates.

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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