Product Details
| abID | ab204709 |
|---|---|
| Cat # +Size | K334-100 |
| Size | 100 assays |
| Detection Method | Fluorescence (Ex/Em = 535/587 nm) |
| Applications | Measurement of HAT activity in Nuclear Extracts Measurement of HAT activity of purified enzyme preparations |
| Features & Benefits | • Simple, rapid & convenient • The assay can detect HAT Activity less than 6 mU in a variety of samples |
| Kit Components | • HAT Assay Buffer • Acetyl CoA (Lyophilized) • H3 Peptide (Lyophilized) • Substrate Mix (Lyophilized) • Developer • PicoProbe™ • CoA Standard (Lyophilized) • Positive Control (HeLa Nuclear Extract) |
| Storage Conditions | -80°C |
| Shipping Conditions | Dry Ice |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Histone Acetyltransferases (HATs) are enzymes that acetylate histone substrates resulting in important regulatory effects on chromatin structure and assembly, and gene transcription. Modifications of these proteins by HATs play an important role in the control of gene expression, and their dysregulation has been linked to cancer, neurodegeneration, and other diseases. BioVision’s HAT Activity Assay Kit utilizes Acetyl CoA and H3 histone peptide as substrates. In this assay, HAT enzyme catalyzes the transfer of acetyl groups from Acetyl-CoA to the histone peptide, thereby generating two products - acetylated peptide and CoA-SH. The CoA-SH reacts with the developer to generate a product that is detected fluorometrically at Ex/Em = 535/587 nm. The assay can detect HAT activity as low as 6 mU in a variety of samples.
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Can DTT interfere with HDAC activity?
According to the literature, DTT can inhibit HDAC enzymes. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2084391/
Can I use RIPA buffer to prepare my samples?
RIPA buffer typically contains SDS which can disrupt the enzyme activity. Therefore it is not recommended with this kit.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Johnstone M et al., (2017) Characterization of the Pro-Inflammatory Cytokine IL-1β on Butyrate Oxidation in Colorectal Cancer Cells, J Cellular Biochem, 2017, 118(6):1614-1621
Stewart et al., Shed Syndecan-1 Translocates to the Nucleus of Cells Delivering Growth Factors and Inhibiting Histone Acetylation: A NOVEL MECHANISM OF TUMOR-HOST CROSS-TALK. J. Biol. Chem., Jan 2015; 290: 941 - 949.
For more citations of this product click here
