Cell Signaling Technology

Product Pathways - Metabolism

PathScan® Phospho-Acetyl-CoA Carboxylase (Ser79) Chemiluminescent Sandwich ELISA Kit #12116

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Kit Includes Volume Solution Color
Phospho-ACC (Ser79) Rabbit Ab Coated Microwells 96 tests
ACC Mouse Detection mAb 5.5 ml Green
Anti-mouse IgG, HRP-linked Antibody 5.5 ml Red
Luminol/Enhancer Solution 3 ml Colorless
Stable Peroxide Buffer 3 ml Colorless
Sealing Tape 2 sheets
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
PathScan® Sandwich ELISA Lysis Buffer (1X) #7018 30 ml Colorless

Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

The PathScan® Phospho-Acetyl-CoA Carboxylase (Ser79) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of acetyl-CoA carboxylase (ACC) protein phosphorylated at Ser79 with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using smaller samples. A Phospho-ACC (Ser79) Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, phospho-ACC protein is captured by the coated antibody. Following extensive washing, an ACC Mouse Detection mAb is added to detect the captured ACC protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-ACC (Ser79) protein.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

PathScan® Phospho-Acetyl-CoA Carboxylase (Ser79) Chemiluminescent Sandwich ELISA Kit recognizes endogenous levels of phospho-ACC (Ser79) in human cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Figure 1. Relationship between protein concentration of lysates from untreated and H2O2-treated Hep G2 cells and immediate light generation with chemiluminescent substrate is shown. Hep G2 cells (80-90% confluent) were treated with H2O2 (10 mM, 10 min) and lysed with PathScan® Sandwich ELISA Lysis Buffer (1X) #7018. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.

Background

Acetyl-CoA carboxylase (ACC) catalyzes the carboxylation of acetyl-CoA to malonyl-CoA (1). It is the key enzyme in the biosynthesis and oxidation of fatty acids (1). In rodents, the 265 kDa ACC1 (ACCα) form is primarily expressed in lipogenic tissues, while 280 kDa ACC2 (ACCβ) is the main isoform in oxidative tissues (1,2). However, in humans, ACC2 is the predominant isoform in both lipogenic and oxidative tissues (1,2). Phosphorylation by AMPK at Ser79 or by PKA at Ser1200 inhibits the enzymatic activity of ACC (3). ACC is a potential target of anti-obesity drugs (4,5).

  1. Castle, J.C. et al. (2009) PLoS One 4, e4369.
  2. Kreuz, S. et al. (2009) Diabetes Metab Res Rev 25, 577-86.
  3. Ha, J. et al. (1994) J Biol Chem 269, 22162-8.
  4. Abu-Elheiga, L. et al. (2001) Science 291, 2613-6.
  5. Levert, K.L. et al. (2002) J Biol Chem 277, 16347-50.

Application References

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Protocols

* Product-specific protocol.

Companion Products

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

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