Goat anti nrp1 c 19

Manufactured by Santa Cruz Biotechnology

Goat anti-NRP1 C-19 is an affinity-purified antibody that recognizes the C-19 epitope of Neuropilin-1 (NRP1). NRP1 is a multi-functional receptor that plays a role in angiogenesis, axon guidance, and cell migration. This antibody can be used for the detection of NRP1 in various applications.

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2 protocols using goat anti nrp1 c 19

Protein Extraction and Western Blotting

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Cells were lysed with lysis buffer (50 mM Tris-HCl, pH 7.4, 5 mM EDTA, 0.1% NP-40, and 250 mM NaCl) in the presence of protease inhibitor cocktail 2 and phosphatase inhibitor cocktail (Sigma-Aldrich). Lungs from P7 mice were lysed in GL35 (Cytoskeleton, Inc). 25 µg of protein was resuspended in Laemmli sample buffer, denatured for 5 min, separated by SDS-PAGE, and transferred to nitrocellulose membrane (Whatman).
Protein extracts were analyzed by SDS-PAGE and transferred to nitrocellulose membrane (Whatman) for immunoblotting with the following primary antibodies: rabbit anti-GAPDH (Abcam), goat anti-NRP1 C-19 (Santa Cruz Biotechnology, Inc.) or AF566 (R&D Systems); mouse anti-ABL1 (BD); rabbit anti-pCRKL (Y207), rabbit anti-pKDR (Y1175) or KDR, rabbit anti-pPXN (Y118), rabbit anti-pAKT (S473) or AKT, or rabbit anti-pERK1/2 or ERK1/2 (Cell Signaling Technology).

Raimondi C., Fantin A., Lampropoulou A., Denti L., Chikh A, & Ruhrberg C. (2014). Imatinib inhibits VEGF-independent angiogenesis by targeting neuropilin 1–dependent ABL1 activation in endothelial cells. The Journal of Experimental Medicine, 211(6), 1167-1183.

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Recombinant Protein and Antibody Analysis

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Recombinant human VEGF₁₆₅ (Ala27-Arg191, no. 293-VE), human Sema3C (Gly21-Ser738, no. 5570-S3), and mouse Sema3C (Gln24-Ser741, no. 1728-S3) were from R&D Systems. Recombinant human IgG-Fc (009-000-008) was purchased from Jackson ImmunoResearch. Antibodies: rat anti-Sema3C (clone 238835) and goat anti-Nrp-1 (AF566) are from R&D Systems; rabbit anti-pY397FAK, rabbit anti-pT180/pY182-p38MAPK (Invitrogen); rabbit anti-FAK (C-20), and goat anti-Nrp-1 (C-19) are from Santa Cruz; mouse anti-p38MAPK (L53F8), rabbit anti-AKT, rabbit anti-pS473AKT, and rabbit anti-cleaved Caspase-3 (Asp175) are from Cell Signaling; pTyr118 paxillin (NEB), rat anti-CD34 (NCL-END; Novocastra); rat anti-CD31 (clone Mec13.3) and rat anti-VE-cadherin are from BD Biosciences; mouse anti-vinculin (V9131) and anti-actin smooth muscle-Cy3 are from Sigma; rabbit anti-GFAP (DAKO), rabbit anti-NG2 (Millipore), rabbit anti-collagen IV (Cosmo Bio).

Yang W.J., Hu J., Uemura A., Tetzlaff F., Augustin H.G, & Fischer A. (2015). Semaphorin-3C signals through Neuropilin-1 and PlexinD1 receptors to inhibit pathological angiogenesis. EMBO Molecular Medicine, 7(10), 1267-1284.

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