Anti-AMPK alpha 1/2 Monoclonal Antibody for WB, IF/ICC, IHC-P, ELISA - Q13131 / P54646

Core Product Specifications and Parameters

Biological Background: AMPK alpha 1/2 Function and Localization

• AMPK catalytic subunit alpha-1 (PRKAA1) acts as a central energy sensor, activating catabolic pathways and inhibiting anabolic processes upon cellular energy stress to restore ATP levels. Related references: PMID:17307971 PMID:17712357 PMID:24563466
• In lipid metabolism, AMPK directly phosphorylates and inactivates acetyl-CoA carboxylase (ACACA/ACACB) and HMG-CoA reductase (HMGCR), thereby suppressing fatty acid and cholesterol synthesis.
• AMPK governs transcriptional reprogramming by phosphorylating key transcription factors and coactivators (e.g., CRTC2, FOXO3, SREBF1, PPARGC1A) and histone H2B, modulating metabolic gene expression. Related references: PMID:11518699 PMID:11554766 PMID:15866171
• Through phosphorylation of RPTOR and TSC2, AMPK inhibits the mTORC1 complex, leading to downregulation of cell growth and proliferation, and promotes autophagy under nutrient deprivation. Related references: PMID:14651849 PMID:18439900 PMID:20160076 PMID:21205641
• Subcellularly, AMPK alpha-1 localizes predominantly to the cytoplasm and nucleus, and is also detected on late endosomes, consistent with its diverse signaling roles.
• The protein is subject to extensive phosphorylation at multiple serine/threonine residues and, based on proteomics identification, may carry N-linked glycosylation.
• AMPK alpha-1 also displays tau-protein kinase activity and can phosphorylate MAPT/TAU in response to amyloid beta, although the in vivo significance remains unclear.
• This antibody recognizes both the alpha-1 catalytic subunit (encoded by PRKAA1, UniProt Q13131) and the highly homologous alpha-2 subunit (encoded by PRKAA2, UniProt P54646), making it a pan-AMPK-alpha tool.

Experimental Guidance and Technical Tips

• The immunogen spans residues 13–300 of human AMPK alpha-1, encompassing the kinase domain. This region is well-conserved across human, mouse, and rat, supporting the reported broad species cross-reactivity.
• For Western blotting, expect a predominant band at approximately 64 kDa. Endogenous AMPK alpha-1 is widely expressed; consider using lysates from cell lines such as HeLa, HEK293T, or tissues like liver and skeletal muscle as positive controls.
• In immunofluorescence and immunohistochemistry, both cytoplasmic and nuclear signals are anticipated. Subcellular localization may shift under specific stress conditions (e.g., glucose deprivation, AMPK activators), so fixation and permeabilization protocols should be optimized for the target compartment.
• For ELISA-based assays, the monoclonal nature of this antibody ensures batch-to-batch consistency; however, users should validate optimal coating and detection conditions in their specific assay system.

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