Histone Acetyltransferase Activity Assay Kit (Colorimetric) (ab65352)
Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Nuclear Extracts, Purified protein, Tissue Lysate
Overview
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Product name
Histone Acetyltransferase Activity Assay Kit (Colorimetric)
See all Histone acetyltransferase kits -
Detection method
Colorimetric -
Sample type
Nuclear Extracts, Purified protein, Tissue Lysate -
Assay type
Enzyme activity -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
Abcam's Histone Acetyltransferase Activity Assay Kit offers a convenient, nonradioactive system for a rapid and sensitive detection of HAT activity in mammalian samples. The kit utilizes active Nuclear Extract (NE) as a positive control and acetyl-CoA as a cofactor. Acetylation of peptide substrate by active HAT releases the free form of CoA which then serves as an essential coenzyme for producing NADH. NADH can easily be detected spectrophotometrically upon reacting with a soluble tetrazolium dye. The detection can be continuous and suitable for kinetic studies. The kit provides a simple, straightforward protocol for a complete assay.
Visit our FAQs page for tips and troubleshooting.HAT assay protocol summary:
- add samples to wells
- add HAT assay mix
- incubate for 1-4 hrs
- analyze with microplate reader -
Notes
Histone acetyltransferases (HATs) have been implicated to play a crucial role in various cellular functions, such as gene transcription, differentiation, and proliferation.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components Identifier 100 tests 2X HAT Assay Buffer Amber 1 x 7.5ml HAT Reconstitution Buffer Clear 1 x 1.8ml HAT Substrate I Blue 1 vial HAT Substrate II Red 1 vial NADH Generating Enzyme Green 1 x 800µl Nuclear Extract (4 mg/ml ) Violet 1 x 50µl -
Research areas
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Relevance
Histone acetyltransferases (HAT) are enzymes that acetylate conserved lysine amino acids on histone proteins. They have been implicated to play a crucial role in various cellular functions, such as gene transcription, differentiation, and proliferation. -
Cellular localization
Cytoplasmic and Nuclear. Note=Nuclear in S-phase cells and cytoplasmic. -
Alternative names
- HAT
- HAT1
- Histone acetyltransferase 1
see all
Images
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Functional studies - ab65352 Image from Goriki A et al., PloS Biol 12(4), Figure S11. doi:10.1371/journal.pbio.1001839. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
HAT activity in NIH3T3 cells, which were transfected with the desired plasmids by using Lipofectamine 2000. After 24 hours from transfection, the nuclear protein was extracted and the Histone acetyltransferase activity was measured by using ab65352.
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Functional studies - ab65352 Image from Jha et al., PLoS One 12(5), fig 2C. doi: 10.1371/journal.pone.0177372. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Histone acetylation activity assay for L.donovani cells over-expressing HAT1, HAT2, HAT3 and HAT4. The activities were compared to wild-type (WT) cells and pLPneo2 (vector) only transfected cells. The relative activity was determined using Histone Acetyltransferase acitvity assay kit (ab65352).
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Functional Studies - Histone Acetyltransferase Activity Assay Kit (Colorimetric) (ab65352)Analysis of Histone Acetyltransferase Activity in HeLa Nuclear Extract using ab65352. HeLa nuclear extract in various amounts was incubated with HAT substrate. Activity was analyzed in a micro plate reader at 440 nm according to the kit instructions.
