Anti igf1 receptor β

Immunoblot assays were performed as described previously [62 (link)]. Primary antibodies were used as followed: anti-E-cadherin (610182, BD, San Jose, CA, USA), anti-α-catenin (610193, BD), anti-N-cadherin (610920, BD), anti-Vimentin (MS-129-P0, Thermo Scientific, Cheshire, UK), anti-Twist (sc-15393, Santa Cruz, Santa Cruz, CA, USA), anti-Snail (3895, Cell Signaling, Danvers, MA, USA), anti-Slug (AP2053a, Abgent, San Diego, CA, USA), anti-MMP2 (#4022, Cell Signaling), anti-MMP9 (#2551-1, Epitomics, Burlingame, CA, USA), anti-phospho-mTOR (Ser2448) (#5536, Cell Signaling), anti-mTOR (#2972, Cell Signaling), anti-phospho-IGF1 Receptor β (Tyr980) (#4568, Cell Signaling), anti-IGF1 Receptor β (#3018, Cell Signaling), anti-p21 (#2946, Cell Signaling), anti-p27 (#2552, Cell Signaling), anti-cyclin D1 (#2926, Cell Signaling), anti-cyclin E (sc-247, Santa Cruz), anti-β-actin (sc-1615, Santa Cruz) and anti-α-tubulin (MS-581-P0, Thermo Scientific). Protein levels were determined by measuring the intensity of bands on the blots using Image J (National Institutes of Health, Bethesda, Maryland, USA). Protein levels were normalized against an internal control β-actin or α-tubulin. The ratio was determined by dividing the normalized protein levels in expressing cells with that in control cells. The mean of ratio was obtained by averaging the ratios from several independent blots.

Anti Insulin Receptor β (4B8) (#3025)(1:1000 working dilution); Anti IGF-1 Receptor β (#3027) (1:1000); Anti Akt2 (D6G4) (#3063) (1:1000); Anti phospho-IGF-1 Receptor β (Tyr1135/1136) / Insulin Receptor β (Tyr1150/1151) (19H7) (1:500); Anti Phospho-Akt (Ser473) (193H12) (#4058) (1:1000); Anti phospho-PDGFR-α (Tyr754) (#2992) (1:1000); Anti PDGFR-α (#3174) (1:1000) were supplied by Cell Signalling Technology^®. Pan 14-3-3 (K-19) (#sc-629) (1:1000) was from Santa Cruz Biotechnology Inc. Anti phospho-IRS1 (Tyr 608) (#09-432) (1:1000) and anti IRS1 (#06-248) (1:500) were supplied by Millipore™. Secondary antibody; Goat anti-Rabbit IgG (H + L) Cross-Adsorbed Secondary Antibody, HRP (#A16104) (1:4000) was supplied by Invitrogen.