TAE684, crizotinib (PF-02341066), LDK378 (ceritinib), lorlatinib, dasatinib, and KD025 were purchased from Selleck Chemicals (Selleckchem). LY294002 and U0126 were purchased from Cell Signaling Technology. The stock solutions were prepared in DMSO and stored at −20°C. IGF-1 was purchased from PeproTech, and insulin was purchased from Sigma-Aldrich. The following antibodies were used: rabbit anti–phospho-ALK (Tyr1604), mouse anti-ALK (31F12), rabbit anti-ALK (C26G7), rabbit anti–phospho-Akt (Ser473) and anti-Akt, mouse anti–phospho-ERK1/2 (Thr202/Tyr204), rabbit anti-ERK1/2, rabbit anti–phospho-FoxO3a (Ser253) (D18H8) and anti-FoxO3a (D19A7), rabbit anti-PI3K p110α and anti-PI3K p85, rabbit anti-IRS1, rabbit anti-IRS2 (L1326), mouse anti-phosphotyrosine (P-Tyr-100), rabbit anti–cleaved caspase-3, rabbit anti–phospho–IGF-1Rβ (Tyr1131)/insulin receptor β (Tyr1146), rabbit anti–insulin receptor β (4B8), and rabbit anti–IGF-1Rβ (D23H3) (Cell Signaling Technology); rabbit anti-IRS2 for immunoprecipitation, mouse anti-GAPDH, anti-SHP2 [M163], and anti-Shc (Abcam); mouse anti-Grb2 (BD Biosciences, San Jose, CA); mouse anti-vinculin (Sigma-Aldrich); mouse anti-phosphotyrosine (4G10) (Millipore, Bedford, MA); and goat anti-rabbit horseradish peroxidase (HRP)–conjugated secondary antibody and goat anti-mouse HRP-conjugated secondary antibody (Jackson ImmunoResearch Laboratories). For phosphoproteomics, anti-phosphotyrosine immunoaffinity beads (P-Tyr-100 and P-Tyr-1000) were purchased from Cell Signaling Technology.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.