Anti-Nuclear Pore Complex Proteins Antibody, clone 414

Nuclear pore glycoprotein p62 (UniProt P17955; also known as 62 kDa nucleoporin, Nucleoporin Nup62) is encoded by the Nup62 gene (Gene ID 65274) in rat species. The nuclear pore complex (NPC) acts as a passageway for nucleocytoplasmic transport. Soluble cargo-protein complexes navigate through the pore by binding to phenylalanine-glycine (FG)-repeat proteins attached to the channel walls. The Nup62 complex is located in the center of the NPC and contains the FG-repeat proteins Nup62, Nup54, and Nup58 associated with each other via conserved coiled-coil segments in a 1:1:1 stiochiometric ratio. The nuclear envelope breaks down during prometaphase. The disassembled nucleoporin subcomplexes act as critical regulators of various mitotic events to ensure proper chromosome segregation. Specifically, Nup62 is reported to localize to centrosomes/spindle poles, where it associates with the centrosomal proteins gamma-tubulin and hSAS-6. Nup62 is involved in the recruitment of critical proteins to the centrosome and its depletion results in mislocalization of several centrosomal components.

Clone 414 immunostained rat liver cell nuclear rim and yeast cell nuclear envelope (Aris, J.P., and Blobel, G. (1989). J. Cell Biol. 108(6):2059-2067; Davis, L.I., and Blobel, G. (1986). Cell. 45(5):699-709).

Triton X-100 treated rat liver nuclei.

Anti-Nuclear Pore Complex Proteins Antibody, clone 414 is an antibody against Nuclear Pore Complex Proteins for use in Immunocytochemistry, Immunoprecipitation, Western Blotting, Electron Microscopy.

Immunocytochemistry Analysis: A 1:200 dilution from a representative lot immunostained NIH/3T3 cell nuclear rim.
Immunocytochemistry Analysis: A representative lot immunostained yeast nuclear envelope with a punctate and patchy pattern (Aris, J.P., and Blobel, G. (1989). J. Cell Biol. 108(6):2059-2067).
Immunocytochemistry Analysis: Representative lots detected a punctate staining pattern of Nup62 at the nuclear rim by fluorescent immunocytochemistry using 2% formaldehyde-fixed, methanol-permeabilized Buffalo rat liver (BRL) cells (Davis, L.I., and Blobel, G. (1987). Proc. Natl. Acad. Sci. U. S. A. 84(21):7552-755; Davis, L.I., and Blobel, G. (1986). Cell. 45(5):699-709).
Electron Microscopy: A representative lot immunostained yeast nuclear envelope using 3% paraformaldehyde/0.2% glutaraldehyde-fixed yeast nuclei LR White sections (Aris, J.P., and Blobel, G. (1989). J. Cell Biol. 108(6):2059-2067).
Electron Microscopy: A representative lot immunostained the pore complexes in thin sections of isolated rat liver nuclei extracted with 2% Triton X-100 and fixed with 0.05% glutaraldehyde (Davis, L.I., and Blobel, G. (1986). Cell. 45(5):699-709).
Immunoprecipitation Analysis: A representative lot immunoprecipited a ~100 kDa (p110) and a ~95 kDa (p95) protein species from yeast nuclear extract. An additional ~55 kDa protein was immunoprecipitated by clone 414 using yeast cytosolic fraction or whole cell lysate (Aris, J.P., and Blobel, G. (1989). J. Cell Biol. 108(6):2059-2067).
Immunoprecipitation Analysis: A representative lot immunoprecipitated three major GlcNAcylated prtoein species of 62, 175, and 270 kDa from SDS-solubilized pore complex-lamina extract of Buffalo rat liver (BRL) cell nuclei preparation that had been labeled with UDP-[3H]Gal by galactosyltransferase (Davis, L.I., and Blobel, G. (1987). Proc. Natl. Acad. Sci. U. S. A. 84(21):7552-755).
Immunoprecipitation Analysis: A representative lot immunoprecipitated GlcNAcylated 62 kDa protein (p62) from the SDS-solubilized pore complex-lamina extract, as well as a less glycosylated p61 cytoplasmic form from the postmitochondrial supernatant of Buffalo rat liver (BRL) cells (7552-755; Davis, L.I., and Blobel, G. (1986). Cell. 45(5):699-709).
Western Blotting Analysis: A representative lot detected a ~100 kDa (p110) and a ~95 kDa (p95) immunoreactive bands in yeast nuclear extract (Aris, J.P., and Blobel, G. (1989). J. Cell Biol. 108(6):2059-2067).
Western Blotting Analysis: A representative lot detected a ~62 kDa (p62) and a ~200 kDa target bands associated with nuclear pore complex-lamina of rat liver nuclei preparation even following sequential nucleaases, Triton X-100, and 140 mM NaCl treatments (Davis, L.I., and Blobel, G. (1986). Cell. 45(5):699-709).

Research Category
Signaling

Research Sub Category
Chromatin Biology

Evaluated by Immunocytochemistry in HeLa cells.

Immunocytochemistry Analysis: A 1:200 dilution of this antibody immunostained HeLa cell nuclear rim.

53.40 kDa (Rat Nup62) calculated. ~62/175/270 kDa (Rat liver nuclear extract) and ~95/100 kDa (Yeast nuclear extract) reported (Aris, J.P., and Blobel, G. (1989). J. Cell Biol. 108(6):2059-2067; Davis, L.I., and Blobel, G. (1986). Cell. 45(5):699-709). Uncharacterized band(s) may appear in some lysates.

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to lot specific datasheet.

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