Lkb1 27d10

Manufactured by Cell Signaling Technology

LKB1 (27D10) is an antibody product from Cell Signaling Technology. It recognizes the LKB1 protein, which is a serine/threonine protein kinase that plays a key role in the regulation of cellular energy metabolism.

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2 protocols using lkb1 27d10

Western Blot Analysis of Cellular Signaling

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Equal amounts of protein were mixed with SDS sample buffer and separated on SDS-PAGE before Western blot analysis. Lysates were homogenized and run on pre-cast SDS-PAGE gels (BioRad, Hercules, CA). The primary antibodies used were: LKB1 (27D10), AMPKα (23A3), phospho-AMPKα (Thr172) (40H9), p70 S6 Kinase (49D7), phospho-p70 S6 Kinase (Thr389), 4E-BP1 (53H11), Phospho-4E-BP1 (Thr37/46) (236B4), Akt (C67E7), phospho-Akt (Ser473) (D9E), Akt2 (D6G4), phospho-Akt2 (Ser474) (D3H2), CDK2 (78B2), Cyclin D1, phospho-Rb (Ser807/811) (D20B12), p27 Kip1 (D69C12), b-actin (all from Cell Signaling Technology, Danvers, MA), Cyclin A (H-432), and GAPDH (FL-335) (all from Santa Cruz Biotechnology, Dallas, TX). Quantification of the Western blot data were performed by measuring the intensity of the hybridization signals using ImageJ analysis program (National Institute of Health, Bethesda, MD).

Shukuya T., Yamada T., Koenig M.J., Xu J., Okimoto T., Li F., Amann J.M, & Carbone D.P. (2019). The effect of LKB1 activity on the sensitivity to PI3K/mTOR inhibition in non-small cell lung cancer. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 14(6), 1061-1076.

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Comprehensive Immunoblotting Protocol for Cellular Signaling

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Antibodies obtained from Cell Signaling Technologies: Anti- phospho-AMPK(T172), pan AMPKα, phospho-ACC(S79), phospho-EGFR(Tyr1173), phospho-S6(S235/236), phospho- S6(S240/244), phospho-4EBP1(T70), 4EBP1(53H11), phospho-Erk1/2 (Thr202/Tyr204), Erk1/2, phospho-AKT(S473), AKT, cleaved caspase 3, LKB1(27D10), PTEN(138G6), BIM (C34C5) and phospho-Histone H3. Antibodies obtained from Santa Cruz: EGFR, S6. Anti-phospho-EGFR(Tyr1173) and anti-AMPK-a1 antibodies were obtained from Millipore. Anti- SLC22A1 (OCT1) and Ki-67 antibodies were obtained from Abcam. Anti-SLC22A2 (OCT2) antibody was obtained from Sigma-Aldrich. Anti-p53 antibody was obtained from Calbiochem.

Lau Y.K., Du X., Reyannavar V., Hopkins B., Shaw J., Bessler E., Thomas T., Pires M.M., Keniry M., Parsons R.E., Cremers S., Szabolcs M, & Maurer M.A. (2014). Metformin and erlotinib synergize to inhibit basal breast cancer. Oncotarget, 5(21), 10503-10517.

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