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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.
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Histone acetyltransferases (HATs) have been implicated to play a crucial role in various cellular functions, such as gene transcription, differentiation, and proliferation.
|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||Purple||1 vial|
|NADH Generating Enzyme||Green||1 x 800µl|
|Nuclear Extract (4 mg/ml )||Red||1 x 50µl|
Our Abpromise guarantee covers the use of ab65352 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent dilution.|
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.
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).