Acetyl CoA Assay Kit (ab87546)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 20 min
- Sample type: Cell Lysate, Tissue Lysate
- Sensitivity: 0.01 nmol/well
Overview
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Product name
Acetyl CoA Assay Kit -
Detection method
Fluorescent -
Sample type
Cell Lysate, Tissue Lysate -
Assay type
Quantitative -
Sensitivity
> 0.01 nmol/well -
Range
0.01 nmol/well - 1 nmol/well -
Assay time
0h 20m -
Product overview
Acetyl CoA Assay Kit ab87546 is a highly sensitive assay for quantifying Acetyl CoA level in biological samples.
In the Acetyl CoA assay protocol, free CoA is quenched then Acetyl CoA is converted to CoA. The CoA is reacted to form NADH which interacts with a probe to generate fluorescence (Ex=535/Em=587 nm).
The assay can detect 10 to 1000 pmol of Acetyl CoA (with detection limit ~0.4 µM).
Acetyl CoA assay protocol summary:
- add samples and standards to wells
- add CoA quencher to wells to remove background from free CoA and succ-CoA and incubate at room temp for 5 min
- add quencher remover and incubate at room temp for 5 min
- add reaction mix and incubate for 10 min at 37ºC
- analyze with microplate reader -
Notes
This product was previously called PicoProbe Acetyl CoA Assay Kit.
Acetyl CoA is a central molecule of metabolism. It carries acetate, used in the build-up and breakdown of larger molecules.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components Identifier 100 tests Acetyl CoA Assay Buffer WM 1 x 25ml Acetyl CoA Enzyme Mix Purple 1 x 500µl Acetyl CoA Standard (1 µmol) (Lyophilised) Yellow 1 vial Acetyl CoA Substrate Mix (Lyophilised) Red 1 vial CoA Quencher Orange 1 x 1ml Conversion Enzyme Green 1 x 100µl PicoProbe™ (in DMSO) Blue 1 x 200µl Quench Remover (Lyophilised) Clear 1 vial -
Research areas
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipid metabolism
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
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Relevance
Acetyl CoA (AcCoA) is the metabolic intermediate which transfers carbon atoms to the TCA cycle for energy production and is the link between fat and carbohydrate metabolism. When fatty acid levels are high, AcCoA levels rise beyond the energy requirements of the organism and the excess is converted to the ketone bodies acetone and ß-hydroxybutyrate. AcCoA also contributes acetate to histone acetylation and isoprenoid synthesis. -
Alternative names
- AcCoA
Images
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Functional Studies - PicoProbe™ Acetyl CoA Assay Kit (ab87546) Image from Zhang Y et al., PLoS One. 2016;11(3):e0150454. Fig 5(A).; doi: 10.1371/journal.pone.0150454. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Induction of acetyl-CoA in mitochondria by palmitate. Mitochondria were isolated from cells after palmitate treatment (300 μM) at 1, 4 and 16 h, and then used in the concentration assay of acetyl-CoA.
The acetyl-CoA content was determined immediately after isolation of the mitochondria using ab87546. Briefly, acetyl-CoA standard curve was made in the range of 0–100 pM and the correlation coefficient was 0.990 or higher. Protein was removed in the sample using the perchloric acid protocol and the supernatant was neutralized with 3 M KHCO3. The CoASH Quencher and Quencher remover were added into the sample to correct the background generated by free CoASH and succ-CoA. The sample was then diluted with the reaction mix, and the fluorescence signal was measured at Ex/Em = 535/589 nm with Spectra max Gemini XPS (Molecular Devices, Sunnyvale, CA). The relative acetyl-CoA concentration was normalized with the mitochondrial protein.
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Functional Studies - PicoProbe™ Acetyl CoA Assay Kit (ab87546)Examples of standard curves generated following the ab87546 kit protocol.