Immunoblotting Analysis of Metabolic Regulators

Lysates from cells or xenografts were prepared in RIPA buffer supplemented with protease/phosphatase inhibitor cocktail (Roche Diagnostics, Indianapolis, IN). Proteins were analyzed by immunoblotting as previously described.^{58 (link)} The antibodies for ALK, β-tubulin, phospho-ALK^Y1604, HK1, HK2, PFK1, GAPDH, PKM2, LDHA, PGAM1, Enolase-1, AKT, phospho-AKT^S473 and phospho-4E-BP1^T37/46 were purchased from Cell Signaling. The antibodies for GLUT1 and GLUT3 were obtained from Abcam (Cambridge, MA). The HIF1α antibody was from BD Biosciences (San Jose, CA).

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Ma Y., Yu C., Mohamed E.M., Shao H., Wang L., Sundaresan G., Zweit J., Idowu M, & Fang X. (2016). A causal link from ALK to hexokinase II overexpression and hyperactive glycolysis in EML4-ALK-positive lung cancer. Oncogene, 35(47), 6132-6142.

Publication 2016

protease/phosphatase inhibitor cocktail β-tubulin phospho-ALKY1604 GAPDH PGAM1 Enolase-1 phospho-AKTS473 phospho-4E-BP1T37/46 antibodies for GLUT1 and GLUT3 HIF1α antibody

Antibodies Antibody Buffer Cells Diagnostics Enolase 1 Gapdh Glut1 Glut3 Hif1α Ldha Pgam1 Phosphatase Protease inhibitor Proteins Ripa Tubulin Xenografts

Corresponding Organization :

Other organizations : Virginia Commonwealth University, Wuhan University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein levels of ALK, β-tubulin, phospho-ALK^Y1604, HK1, HK2, PFK1, GAPDH, PKM2, LDHA, PGAM1, Enolase-1, AKT, phospho-AKT^S473, phospho-4E-BP1^T37/46, GLUT1, GLUT3, and HIF1α

control variables

RIPA buffer supplemented with protease/phosphatase inhibitor cocktail used for cell/xenograft lysate preparation

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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