Cell Signaling Technology

Product Pathways - Metabolism

AMPK Subunit Antibody Sampler Kit #9839

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
AMPKα1 Antibody #2795 40 µl W H Mk (Pg) 62 Rabbit
AMPKα2 Antibody #2757 40 µl W IP H Mk 62 Rabbit
AMPKβ1 (71C10) Rabbit mAb #4178 40 µl W H M Hm Mk Pg 38 Rabbit IgG
AMPKβ2 Antibody #4148 40 µl W IP H M R Mk 30 Rabbit
AMPKγ1 Antibody #4187 40 µl W H Mk 37 Rabbit
AMPKγ2 Antibody #2536 40 µl W H M R Mk B 75 Rabbit
AMPKγ3 Antibody #2550 40 µl W H 54 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine  Pg=Pig
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each of the antibodies in the AMPK Subunit Antibody Sampler Kit detects endogenous levels of the specified AMPK protein. Antibodies do not cross-react with related AMPK subunit proteins.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 (human), NBT-II (rat), and Neuro-2A (mouse) cells using AMPKγ2 Antibody #2536.

Western Blotting

Western Blotting

Western blot analysis of extracts from RD cells using AMPKγ3 Antibody #2550.

Western Blotting

Western Blotting

Western blot analysis of extracts from HEK293 and COS cells using AMPKα2 Antibody #2757.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, HT29 and COS cells, using AMPKα1 Antibody #2795.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293, C6, L-929 and COS cells using AMPKβ2 Antibody #4148.

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12, COS and H-4-III-E cells using AMPKβ1 (71C10) Rabbit mAb #4178.


Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat and K-562 cells using AMPKγ1 Antibody #4187.

Description

The AMPK Subunit Antibody Sampler Kit provides an economical means to investigate the role played by all AMPK subunits in cellular energy homeostasis. The kit contains enough primary and secondary antibodies to perform four Western blots with each antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu519 near the carboxy terminus of human AMPKα1,corresponding to residues surrounding Ser500 of human AMPKα2, near the amino terminus of human AMPKγ1, surrounding Ser60 of human AMPKγ2, and corresponding to the sequences of human AMPKβ2 and AMPKγ3. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues surrounding Val176 of human AMPKβ1.

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 the 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, Ser108, and Ser182 (6,7). Phosphorylation at Ser108 of the β1 subunit seems to be required for the activation of AMPK enzyme, while phosphorylation at Ser24/25 and Ser182 affects AMPK localization (7). Several mutations in AMPKγ subunits have been identified, most of which are located in the putative AMP/ATP binding sites (CBS or Bateman domains). Mutations at these sites lead to reduction of AMPK activity and cause glycogen accumulation in heart or skeletal muscle (1,2). 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).

  1. Hardie, D.G. (2004) J Cell Sci 117, 5479-87.
  2. Carling, D. (2004) Trends Biochem Sci 29, 18-24.
  3. Hawley, S.A. et al. (1996) J Biol Chem 271, 27879-87.
  4. Lizcano, J.M. et al. (2004) EMBO J 23, 833-43.
  5. Shaw, R.J. et al. (2004) Proc Natl Acad Sci USA 101, 3329-35.
  6. Woods, A. et al. (2003) J Biol Chem 278, 28434-42.
  7. Warden, S.M. et al. (2001) Biochem J 354, 275-83.

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