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2 protocols using thr202 tyr204 erk1 2

1

Evaluating IGF-1R Signaling Pathways

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Total IGF-1R, IRS-1, p85, ERK1/2 and Akt and phosphorylated (Tyr1135/1136) IGF-1R, (Tyr458) p85, (Thr202/Tyr204) ERK1/2, (Ser473) Akt, and GAPDH antibody (rabbit) primary antibodies were obtained from Cell Signaling (Danvers, MA). Secondary goat anti-rabbit IgG antibodies (IRDye 800CW) were purchased from LI-COR Biotechnology (Lincoln, NE). AG 1024, a specific IGF-1R phosphorylation inhibitor, was purchased from Selleck Chemicals (Houston, TX). IGF-1R siRNA was obtained from Cell Signaling (Danvers, MA). D-mannitol and glucose were purchased from Sigma (St. Louis, MO). KZ-41 (Fig 1) was synthesized in Dr. Duane Miller’s laboratory and verified to be >96% pure by nuclear magnetic resonance spectroscopy [17 (link)].
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2

Comprehensive Signaling Pathway Profiling

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(Ser473)-Akt, (Ser235/236)-S6, (Ser32/36)-IκB, (Thr202/Tyr204)-Erk1/2, Akt, S6, Erk1/2, IκB, p65, CD79A, Mcl1 and GAPDH antibodies were purchased from Cell Signaling, Danvers, MA, USA. Anti-NOXA was obtained from Calbiochem, La Jolla, CA, USA. Bez235, Ibrutinib, PD0325901, MG132, Bortezomib, LY294002 and BKM120 were obtained from Selleckchem, Houston, TX, USA. Temsirolimus, Orlistat and cycloheximide were purchased from Sigma, Munich, Germany. MLN4924 was obtained from ChemieTek, Indianapolis, IN, USA.
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