Ampk α 23a3

Manufactured by Cell Signaling Technology

Sourced in United States

AMPK-α (23A3) is a primary antibody produced by Cell Signaling Technology. It recognizes the AMPK-α subunit of the AMP-activated protein kinase (AMPK) enzyme, which is a cellular energy sensor that plays a critical role in regulating metabolism. This antibody can be used for applications such as Western blotting to detect the AMPK-α subunit.

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Lab products found in correlation

3 protocols using ampk α 23a3

Comprehensive Antibody Detection Protocol

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Anti-tubulin βIII antibody (clone Tuj1; R&D Systems), at 1:20,000 for Western blot, 1:1,000 for immunoistochemistry (IHC) staining. P-AMPK thr172: phospho-AMPK-α (Thr172) (40H9) Cell Signaling, at 1:1,000. AMPK: AMPK-α (23A3) Cell Signaling, at 1:1,000. P-ACC ser 79: phospho-ACC (Ser79) Cell Signaling, at 1:1,000. ACC: anti-acetyl coenzyme A carboxylase antibody [EP687Y], AB-ab45174 Abcam, at 1:2,000. P-ULK ser555: phospho-ULK1 (Ser555) (D1H4) Cell Signaling, at 1:1,000. Caspase-3-cleaved: cleaved caspase-3 (Asp175) Cell Signaling, at 1:200. Islet antibody: Developmental Studies Hybridoma bank 39.3F7, at 1:200. Tom20: Santa Cruz Tom20 (FL-145): sc 11415, at 1:1,000 for immunofluorescence (STORM analysis) and TOM20 (D8T4N) Cell Signaling at 1:1,000 for immunoblot. β-Actin: β-Actin (13E5) Cell Signaling, at 1:20,000. Efhd1 antibody was a kind gift from the laboratory of Yasuhiro Tomooka (Tokyo University of Science), at 1:1,000. Anti-neurofilament antibody 2H3, Developmental Studies Hybridoma bank, at 1:200.
Antimouse and antirabbit antibodies conjugated with Alexa 549, Alexa 488, or Alexa 647 fluorophores were used at 1:200 (Jackson ImmunoResearch Laboratories). Secondary antibodies for Western blotting: goat antimouse IgG-HRP (JIR 155-035) and goat antirabbit IgG-HRP (JIR 111-035) from Jackson, both at 1:5,000.

Ulisse V., Dey S., Rothbard D.E., Zeevi E., Gokhman I., Dadosh T., Minis A, & Yaron A. (2020). Regulation of axonal morphogenesis by the mitochondrial protein Efhd1. Life Science Alliance, 3(7), e202000753.

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Western Blotting Protocol for Protein Analysis

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The detailed protocol for Western blotting has been described previously.19 The indicating antibodies to the following proteins were used in this study: β‐actin (#4967), EGFR (D38B1) (#4267), phosphor‐EGFR (Tyr1068) (#2234), phospho‐Rb (Ser780) (#9307), p21 Waf1/Cip1 (12D1) (#2947), p27 Kip1 (D69C12) (#3686), phosphor‐Histone H2A.X (Ser139/Tyr142) (#5438), AMPKα (23A3) (#2603), and phospho‐AMPKα (Thr172) (#2535) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Kunimasa K., Nagano T., Shimono Y., Dokuni R., Kiriu T., Tokunaga S., Tamura D., Yamamoto M., Tachihara M., Kobayashi K., Satouchi M, & Nishimura Y. (2017). Glucose metabolism‐targeted therapy and withaferin A are effective for epidermal growth factor receptor tyrosine kinase inhibitor‐induced drug‐tolerant persisters. Cancer Science, 108(7), 1368-1377.

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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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