Protein extraction and western blot analysis was performed as previously described [14 (link), 20 (link)]. The following antibodies were used: anti-phospho IGF-1Rβ(Tyr1135/1136)/Insulin Receptorβ(Tyr1150/1151), anti-total Insulin Receptorβ and total IGF-1Rβ (Cell Signaling Technology, Danvers, MA, USA) and anti-β-actin (Sigma Aldrich, St Louis, MO, USA) at concentrations detailed in electronic supplementary material (ESM) Table 1 . Western blots were imaged and quantified by densitometry after incubation with LI-COR IRdye secondary antibodies with a LI-COR scanner and LI-COR Image Studio software (LI-COR, Lincoln, NE, USA).
Anti phospho igf 1rβ
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-phospho IGF-1Rβ is a laboratory reagent that specifically detects the phosphorylated form of the insulin-like growth factor 1 receptor beta subunit (IGF-1Rβ). It is used to monitor the activation status of the IGF-1 signaling pathway in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Merck Group (2 mentions)
Anti akt, by Cell Signaling Technology (2 mentions)
Anti phospho akt, by Cell Signaling Technology (2 mentions)
Insulin receptor β, by Cell Signaling Technology (1 mentions)
Image studio software, by LI COR (1 mentions)
Licor scanner, by LI COR (1 mentions)
Anti egfr, by Merck Group (1 mentions)
Anti phospho mtor, by Cell Signaling Technology (1 mentions)
Anti tsc2, by Cell Signaling Technology (1 mentions)
Anti phospho pten, by Cell Signaling Technology (1 mentions)
Anti irs1, by Cell Signaling Technology (1 mentions)
Anti met, by Merck Group (1 mentions)
Anti phospho met, by Cell Signaling Technology (1 mentions)
Anti phospho pdgfrβ, by Cell Signaling Technology (1 mentions)
Anti pten, by Cell Signaling Technology (1 mentions)
Anti mtor, by Cell Signaling Technology (1 mentions)
Anti phospho 4ebp1, by Cell Signaling Technology (1 mentions)
Anti tsc1, by Cell Signaling Technology (1 mentions)
Anti erk1 2, by Cell Signaling Technology (1 mentions)
Anti α tubulin antibody, by Merck Group (1 mentions)
4 protocols using anti phospho igf 1rβ
1
Protein Extraction and Western Blot Analysis
2
Antibody Sourcing and Characterization
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Anti-NDRG1 antibody was generated as previously described [43 (link)]. Other antibodies were purchased as follows: anti-α-tubulin antibody was from Sigma-aldrich Co; anti-EGFR, anti-phospho-EGFR (Tyr1068), anti-phospho-HER3, anti-Met, anti-phospho-Met, anti-PDGFRβ, anti-phospho-PDGFRβ, anti- IGF-1Rβ, anti-phospho-IGF-1Rβ, anti-ERK1/2, anti-phospho-ERK1/2, anti-AKT, anti-phospho-AKT (Thr308 and Ser473), anti-mTOR, anti-phospho-mTOR (Ser2448 and Ser2481), anti-Raptor, anti-Rictor, anti-S6K, anti-phospho-S6K(Thr389), anti-phospho-S6, anti-PTEN, anti-phospho-PTEN, anti-IRS-1, anti-PDK1, anti-PARP, anti-phospho-PDK1, anti-TSC-1, anti-TSC-2, and anti-phospho-4EBP-1 antibodies were from Cell Signaling Technology (Beverly, MA); anti-HER2 and anti-HER3 antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA); anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was from Trevigen Inc. (Gaithersburg, MD): anti-p27 antibody was from BD Biosciences (San Jose, CA): anti-cleaved PARP antibody was from Promega (Madison, WI): anti-phospho-HER2 antibody was from Millipore (Billerica, MA): anti-β-actin was from Abcam (Cambridge, UK).
3
Pluripotency Marker Expression Analysis
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Primary antibodies included anti-phospho-IGF1R-β (Tyr1135/1136)/INSR-β (Tyr1150/1151), phospho-AKT (Ser473), AKT, phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, and phospho-GSK3α/β (Ser21/9) (Cell Signaling); INSR-β and IGF1R-β (Santa Cruz); GSK3α/β (Millipore); phospho-insulin receptor substrate 1 (IRS1) (Y612) (Life Technologies); IRS1 (BD Biosciences); NANOG and OCT4 (Abcam); and SSEA4 and TRA-1-60 (Millipore).
4
Targeted Inhibitor Screening in Cancer Cells
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PP242 (#CD0258), Ku-0063794 (#CD0274), and PX-866 (#CD0279) were from Chemdea (Ridgewood, NJ, USA). BKM-120 (#S2247), BYL719 (#S2814), MK-2206 (#S1078), NVP-AEW541 (#S1034), sapitinib (#S2192), and erlotinib (#S7786) were from Selleck Chemicals (Houston, TX, USA). Anti-phospho-AKT (#4060), anti-AKT (#2920), and anti phospho-IGF-1Rβ (#3024) antibodies were from Cell Signaling Technology (Danvers, MA, USA). Anti-β-Actin (#A2228) antibody was from Sigma-Aldrich (Buchs, Switzerland). supplemented with 10% FBS and 1% streptomycin/penicillin in a 5% CO2 atmosphere and at 37ºC.
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