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9957
AMPK and ACC Antibody Sampler Kit

AMPK and ACC Antibody Sampler Kit #9957

Western Blotting Image 1

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

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Western Blotting Image 2

Western blot analysis of extracts from HeLa, K-562, C6, and Neuro-2a cells using AMPKα (D5A2) Rabbit mAb.

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Western Blotting Image 3

Western blot analysis of extracts from oligomycin treated C6 cells or C6 cell lysate treated with λ phosphatase, using Phospho-AMPKβ1 (Ser182) Antibody (upper) and AMPKβ1 Antibody #4182 (lower).

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Western Blotting Image 4

Western blot analysis of cell lysates from various cell lines using AMPKβ 1/2(57C12) Rabbit mAb.

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Western Blotting Image 5

Western blot analysis of extracts from SH-SY5Y cells, untreated or treated with Oligomycin #9996 (0.5 μM, 30 min), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb (upper) or Acetyl-CoA Carboxylase (C83B10) Rabbit mAb #3676 (lower). The phospho-specificity of the antibody was verified by λ phosphatase treatment.

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Western Blotting Image 6

Western blot analysis of cell extracts from various cell lines, using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

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Western Blotting Image 7

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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IHC-P (paraffin) Image 8

Immunohistochemical analysis of paraffin-embedded NCI-H228 cell pellets, control (left) or phenformin-treated (right), using Phospho-AMPKalpha (T172) (40H9) Rabbit mAb.

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IHC-P (paraffin) Image 9

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, showing cytoplasmic localization using AMPKβ1/2 (57C12) Rabbit mAb.

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IHC-P (paraffin) Image 10

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb.

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IHC-P (paraffin) Image 11

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb in the presence of control peptide (left) or Acetyl-CoA Carboxylase (C83B10) Blocking Peptide #1062 (right).

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IHC-P (paraffin) Image 12

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using AMPKβ1/2 (57C12) Rabbit mAb in the presence of control peptide (left) or AMPKβ 1/2 Blocking Peptide #1074 (right).

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IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded mouse liver using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb.

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IHC-P (paraffin) Image 14

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

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Flow Cytometry Image 15

Flow cytometric analysis of COS cells, using AMPKβ1/2 (57C12) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IHC-P (paraffin) Image 16

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb.

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IHC-P (paraffin) Image 17

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

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IF-IC Image 18

Confocal immunofluorescent analysis of untreated C2C12 cells labeled with AMPKβ1/2 (57C12) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IHC-P (paraffin) Image 19

Immunohistochemical analysis of paraffin-embedded NCI-H2228 cell pellets, untreated (left) or phenformin-treated (right), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb.

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IHC-P (paraffin) Image 20

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma, using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

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IF-IC Image 21

Confocal immunofluorescent analysis of 293 cells (all nutrient-starved with Krebs-Ringer bicarbonate buffer for 4 hr), starved only (top left), serum-treated (10%, 30 min; top right), H2O2-treated (10 mM, 10 min; bottom left), or λ phosphatase-treated (2 hr; bottom right), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

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Flow Cytometry Image 23

Flow cytometric analysis of 293 cells, using Acetyl-CoA Carboxylase (C83B10) Rb mAb (blue) compared to a nonspecific negative control antibody (red).

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IF-IC Image 24

Confocal immunofluorescent analysis of NIH/3T3 cells labeled with Acetyl-CoA Carboxylase (C83B10) Rabbit mAb (red). Blue pseudocolor=Draq5™ (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-AMPKα (Thr172) (40H9) Rabbit mAb 2535 20 µl
  • WB
  • IP
  • IHC
H M R Hm Mk Dm Sc 62 Rabbit IgG
AMPKα (D5A2) Rabbit mAb 5831 20 µl
  • WB
  • IP
H M R Mk B 62 Rabbit 
Phospho-AMPKβ1 (Ser182) Antibody 4186 20 µl
  • WB
H M R Mk 38 Rabbit 
AMPKβ1/2 (57C12) Rabbit mAb 4150 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Hm Mk 30, 38 Rabbit IgG
Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb 11818 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R 280 Rabbit IgG
Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 3676 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Hm 280 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

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.

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.

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, 5479-87.
  2. Carling, D. (2004) Trends Biochem Sci 29, 18-24.
  3. Hawley, S.A. et al. (1996) J Biol Chem 271, 27879-87.
  4. Ha, J. et al. (1994) J Biol Chem 269, 22162-8.
  5. Lizcano, J.M. et al. (2004) EMBO J 23, 833-43.
  6. Abu-Elheiga, L. et al. (2001) Science 291, 2613-6.
  7. Levert, K.L. et al. (2002) J Biol Chem 277, 16347-50.
  8. Woods, A. et al. (2003) J Biol Chem 278, 28434-42.
  9. Warden, S.M. et al. (2001) Biochem J 354, 275-83.
  10. Shaw, R.J. et al. (2004) Proc Natl Acad Sci USA 101, 3329-35.
  11. Ruderman, N.B. et al. (1999) Am. J. Physiol. 276, E1-E18.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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