Cell Signaling Technology

Product Pathways - Metabolism

AMPK and ACC Antibody Sampler Kit #9957

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-AMPKα (Thr172) (40H9) Rabbit mAb #2535 40 µl W IP IHC-P H M R Hm Mk Dm Sc (C) (Z) (B) (Pg) 62 Rabbit IgG
AMPKα (23A3) Rabbit mAb #2603 40 µl W H M R Mk 62 Rabbit IgG
Phospho-AMPKβ1 (Ser108) Antibody #4181 40 µl W IP H M R Mk 38 Rabbit
AMPKβ1/2 (57C12) Rabbit mAb #4150 40 µl W IHC-P IF-IC F H M R Hm Mk 30, 38 Rabbit IgG
Phospho-Acetyl-CoA Carboxylase (Ser79) Antibody #3661 40 µl W IP IHC-P H M R Mk (C) (B) 280 Rabbit
Acetyl-CoA Carboxylase (C83B10) Rabbit mAb #3676 40 µl W IHC-P IF-IC F H M R Hm 280 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Dm=D. melanogaster  Z=Zebrafish  B=Bovine  Pg=Pig  Sc=S. cerevisiae
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

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 (23A3) Rabbit mAb detects endogenous levels of total AMPKα protein. Phospho-AMPKβ1 (Ser108) Antibody detects endogenous levels of AMPKβ1 only when phosphorylated at Ser108. The antibody may cross-react with phosphorylated AMPKβ2 when phosphorylated at Ser109.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) Antibody detects endogenous levels of ACC only when phosphorylated at Ser79. The antibody recognizes both ACCα and ACCβ. Acetyl-CoA Carboxylase (C83B10) Rabbit mAb detects endogenous levels of all isoforms of acetyl-CoA carboxylase protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12 cells, untreated or oligomycin-treated (0.5 µM), using Phospho-AMPKα (Thr172) (40H9) Rabbit mAb #2535 (upper) or AMPKα Antibody #2532 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cells and tissues using AMPKα (23A3) Rabbit mAb #2603.

Western Blotting

Western Blotting

Western blot analysis of extracts from HEK293 cells, untreated or oligomycin-treated, using Phospho-Acetyl-CoA Carboxylase (Ser79) Antibody #3661.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb #3676.

Western Blotting

Western Blotting

Western blot analysis of cell lysates from various cell types using AMPKβ1/2 (57C12) Rabbit mAb #4150.

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12 cells, untreated (lanes 1,3) or oligomycin-treated (lanes 2,4), using Phospho-AMPKβ1 (Ser108) Antibody #4181 (upper) or AMPKβ1 Antibody #4182 (lower). Cell lysates were treated with λ phosphatase in lanes 3 and 4 to demonstrate the phospho-specificity of Phospho-AMPKβ1 (Ser108) Antibody.


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 four Western blots with each antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser79 of rat Acetyl-CoA Carboxylase and Ser491 of human AMPKα2. Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr172 of human AMPKα, the amino-terminal sequence of human AMPKα, residues surrounding His231 of AMPKβ1 and residues surrounding Ser52 of human Acetyl-CoA Carboxylase α1. 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).

  1. Hardie, D.G. (2004) J. Cell Sci. 117, 5479-5487.
  2. Carling, D. (2004) Trends Biochem. Sci. 29, 18-24.
  3. Hawley, S.A. et al. (1996) J. Biol. Chem. 271, 27879-27887.
  4. Lizcano, J.M. et al. (2004) EMBO J. 23, 833-843.
  5. Shaw, R.J. et al. (2004) Proc. Natl. Acad. Sci. U S A 101, 3329-3335.
  6. Woods, A. et al. (2003) J. Biol. Chem. 278, 28434-28442.
  7. Warden, S.M. et al. (2001) Biochem J. 354, 275-283.
  8. Ruderman, N.B. et al. (1999) Am. J. Physiol. 276, E1-E18.
  9. Ha, J. et al. (1994) J. Biol. Chem. 269, 22162-22168.
  10. Abu-Elheiga, L. et al. (2001) Science 291, 2613-2616.
  11. Levert, K.L. et al. (2002) J. Biol. Chem. 277, 16347-16350.

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

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