Name: AMPK and ACC Antibody Sampler Kit
Price: 538.00 USD
Availability: InStock

Product Information

Kit Usage Information

Protocols

Product Description

The AMPK and ACC Antibody Sampler Kit provides an economical means to investigate energy homeostasis and fatty acid synthesis within the cell. The kit contains primary and secondary antibodies to perform two Western blots with each antibody.

Specificity / Sensitivity

Phospho-AMPKα (Thr172) (40H9) Rabbit mAb detects endogenous AMPKα only when phosphorylated at Thr172. The antibody detects both α1 and α2 isoforms of the catalytic subunit, but does not detect the regulatory β or γ subunits. AMPKα (D5A2) Rabbit mAb detects endogenous levels of total AMPKα protein. Phospho-AMPKβ1 (Ser182) Antibody detects endogenous levels of AMPKβ1 only when phosphorylated at Ser182. AMPKβ1/2 (57C12) Rabbit mAb detects endogenous levels of both total AMPKβ1 and β2 proteins. The antibody does not cross-react with other related proteins. Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb recognizes endogenous levels of acetyl-CoA carboxylase protein only when phosphorylated at Ser79. Acetyl-CoA Carboxylase (C83B10) Rabbit mAb detects endogenous levels of all isoforms of acetyl-CoA carboxylase protein.

Source / Purification

Monoclonal state-specific antibod- ies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr172 of human AMPKα, and Ser79 of human ACC. Total monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg21 of human AMPKα, His233 of human AMPKß1, and Ser523 of human Acetyl-CoA Carboxylase α1. Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser182 of human AMPKß1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

AMP-activated protein kinase (AMPK) is highly conserved from yeast to plants and animals and plays a key role in the regulation of energy homeostasis (1). AMPK is a heterotrimeric complex composed of a catalytic α subunit and regulatory β and γ subunits, each of which is encoded by two or three distinct genes (α1, 2; β1, 2; γ1, 2, 3)(2). The kinase is activated by an elevated AMP/ATP ratio due to cellular and environmental stress, such as heat shock, hypoxia and ischemia (1). The tumor suppressor LKB1, in association with accessory proteins STRAD and MO25, phosphorylates AMPKα at Thr172 in the activation loop and this phosphorylation is required for AMPK activation (3-5). AMPKα is also phosphorylated at Thr258 and Ser485 (for α1; Ser491 for α2). The upstream kinase and biological significance of these phosphorylation events have yet to be elucidated (6). The β1 subunit is post-translationally modified by myristoylation and multi-site phosphorylation including Ser24/25, Ser96, Ser101 and Ser182 (6,7). Phosphorylation at Ser108 of the β1 subunit seems to be required for the activation of AMPK enzyme, while phosphorylation ot Ser24/25 and Ser182 affects AMPK localization (7). Accumulating evidence indicates that AMPK not only regulates the metabolism of fatty acids and glycogen, but also modulates protein synthesis and cell growth through EF2 and TSC2/mTOR pathways, as well as blood flow via eNOS/nNOS (1).
Acetyl-CoA carboxylase (ACC) catalyzes the pivotal step of the fatty acid synthesis pathway. The 265 kDa ACCα is the predominant isoform found in liver, adipocytes and mammary gland, while the 280 kDa ACCβ is the major isoform in skeletal muscle and heart (8). Phosphorylation by AMPK at Ser79 or by PKA at Ser1200 inhibits the enzymatic activity of ACC (9). ACC is a potential target of anti-obesity drugs (10,11).

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.

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AMPK and ACC Antibody Sampler Kit #9957