Acetyltransferase Activity Assay Kit (Fluorometric) (ab204536)
Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Inhibitor compounds, Purified protein
Overview
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Product name
Acetyltransferase Activity Assay Kit (Fluorometric) -
Detection method
Fluorescent -
Sample type
Purified protein, Inhibitor compounds -
Product overview
Acetyltransferase Activity Assay Kit (Fluorometric) (ab204536) is a homogeneous mix-and-read fluorescent assay for the determination of any acetyl-CoA dependent acetyltransferase activity. It is suitable for end-point or kinetic read options, which is ideal for determining mechanism of action, kinetics, and screening candidate compounds. The assay is amendable to HTS and miniaturization.
This assay is a complete kit for the screening of candidate compounds that can alter normal acetyltransferase activity. For use with purified in vitro samples.
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Notes
Acetylation is an important covalent molecular modification. Originally identified as the method by which certain bacteria were able to deactivate anti-microbial compounds, acetylation is now also known as an important partitioning and signaling modification.
Acetyltransferases are enzymes that covalently transfer an acetyl group from a donor molecule (Acetyl CoA) to an acceptor. Acetyl CoA serves as a universal donor while the acceptor varies with the acetyltransferase. Acceptors include histones, kinases, transcription factors, receptors, neurotransmitter precursors like choline and serotonin, and anti-microbial agents like chloramphenicol and fluoroquinones. Acetylation can signal an increase or decrease in activity based on the context of the message. Frequently located at critical junctions in metabolic pathways, Acetyltransferases and their regulation have become attractive therapeutic targets to treat everything from insomnia to cancer.
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Platform
Microplate reader
Properties
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Storage instructions
Please refer to protocols. -
Components 1 x 96 tests Acetyltransferase Positive Control 1 x 200µl Acetyltransferase Reaction Buffer Concentrate 1 x 100µl Black Microtiter Plate 1 unit Foil Plate Sealer 3 units Transferase Assay Buffer Concentrate 1 x 15ml Transferase Detection Solution Concentrate 1 x 150µl -
Research areas
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Cholesterol Metabolism
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipoprotein metabolism
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Sequence similarities
Belongs to the thiolase family. -
Cellular localization
Cytoplasm. - Information by UniProt
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Alternative names
- Acat2
- acetoacetyl CoA thiolase
- acetoacetyl Coenzyme A thiolase
see all
Images
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Chloramphenicol acetyltransferase (CAT) titration in acetyltransferase assay obtained using Abcam'sAcetyltransferase Activity Assay Kit (Fluorometric) (ab204536).
Chloramphenicol acetyltransferase (CAT) was titered in the assay using 100μM of the Substrate chloramphenicol. Serial dilutions of the enzyme were prepared in Transferase Assay Buffer. Mean relative fluorescence was plotted against CAT concentration to generate the following graph.
This is for illustration purposes only.
The investigator must titrate their enzyme / substrate system in the assay.
Based on this titration data, the acetyltransferase concentration of 100nM produces a maximum signal within the detection range of the plate reader, with a signal to noise ratio sufficient for easy detection of altered enzyme activity.
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Example Kinetic Assay Results.100nM CAT was tested with 100μM Chloramphenicol substrate in the kinetic assay format. Mean relative fluorescence was plotted against the stop time interval to generate this graph. Obtained using Abcam's Acetyltransferase Activity Assay Kit (Fluorometric) (ab204536).
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Typical inhibition curve obtained using Abcam's Acetyltransferase Activity Assay Kit (Fluorometric) (ab204536).
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab204536 has been referenced in 5 publications.
- Luo R et al. A novel missense variant in ACAA1 contributes to early-onset Alzheimer's disease, impairs lysosomal function, and facilitates amyloid-ß pathology and cognitive decline. Signal Transduct Target Ther 6:325 (2021). PubMed: 34465723
- Arun KB et al. Acetylation of Isoniazid Is a Novel Mechanism of Isoniazid Resistance in Mycobacterium tuberculosis. Antimicrob Agents Chemother 65:N/A (2020). PubMed: 33106268
- Ghelani H et al. Atorvastatin Improves Hepatic Lipid Metabolism and Protects Renal Damage in Adenine-Induced Chronic Kidney Disease in Sprague-Dawley Rats. Biomed Res Int 2019:8714363 (2019). PubMed: 31828139
- Counihan JL et al. Chemoproteomic Profiling of Acetanilide Herbicides Reveals Their Role in Inhibiting Fatty Acid Oxidation. ACS Chem Biol 12:635-642 (2017). PubMed: 28094496
- Ford B et al. Mapping Proteome-wide Targets of Glyphosate in Mice. Cell Chem Biol 24:133-140 (2017). PubMed: 28132892