Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate (12 x 8 well strips)
产品名称HDAC Activity Assay试剂盒(Fluorometric)
参阅全部 HDAC 试剂盒
实验步骤Multiple steps standard assay
Inhibition of histone deacetylase (HDAC) has been implicated in the modulation of transcription and induction of apoptosis or differentiation in cancer cells.
However, screening of compounds for HDAC inhibition has been difficult due to the lack of convenient tools for analyzing HDAC activity.
The new Fluorometric HDAC Activity Assay Kit provides a fast and fluorescence-based method that eliminates radioactivity, extractions, or chromatography, as used in traditional assays.
The new procedure requires only two easy steps, both performed on the same microtiter plate.
First, the HDAC fluorometric substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (e.g., HeLa nuclear extract or others).
Deacetylation of the substrate sensitizes the substrate, so that, in the second step, treatment with the Lysine Developer produces a fluorophore.
The fluorophore can be easily analyzed using a fluorescence plate reader or a fluorometer.
The assay is well suited for high throughput screening applications.
Read the entire protocol before beginning the procedure.
The HeLa extract should be refrozen immediately at –70°C after each use to avoid loss of activity.
If positive and negative controls are designed, the kit provides sufficient reagents for 5 positive control assays with the HeLa Nuclear Extract and 5 negative control assays with the HDAC Inhibitor, Trichostatin A.
平台Microplate (12 x 8 well strips)
存放说明Store at -80°C. Please refer to protocols.
组件 标识符 96 tests 10X HDAC Assay Buffer Green cap 1 x 1ml Deacetylated Standard (4 mM) Yellow cap 1 x 20µl HDAC Inhibitor (Trichostatin A, 1mM) Blue Cap 1 x 10µl HDAC Substrate Amber cap 1 x 500µl HeLa Nuclear Extract (5 mg/ml) Red cap 1 x 10µl Lysine Developer Orange cap 1 x 1ml
功能Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Deacetylates SP proteins, SP1 and SP3, and regulates their function. Component of the BRG1-RB1-HDAC1 complex, which negatively regulates the CREST-mediated transcription in resting neurons. Upon calcium stimulation, HDAC1 is released from the complex and CREBBP is recruited, which facilitates transcriptional activation. Deacetylates TSHZ3 and regulates its transcriptional repressor activity. Deacetylates 'Lys-310' in RELA and thereby inhibits the transcriptional activity of NF-kappa-B. Component a RCOR/GFI/KDM1A/HDAC complex that suppresses, via histone deacetylase (HDAC) recruitment, a number of genes implicated in multilineage blood cell development.
组织特异性Ubiquitous, with higher levels in heart, pancreas and testis, and lower levels in kidney and brain.
序列相似性Belongs to the histone deacetylase family. HD type 1 subfamily.
翻译后修饰Sumoylated on Lys-444 and Lys-476; which promotes enzymatic activity. Desumoylated by SENP1.
Phosphorylation on Ser-421 and Ser-423 promotes enzymatic activity and interactions with NuRD and SIN3 complexes. Phosphorylated by CDK5.
Ubiquitinated by CHFR, leading to its degradation by the proteasome (By similarity). Ubiquitinated by KCTD11, leading to proteasomal degradation.
- Information by UniProt
- Histone deacetylase 1
ab1438 被引用在 5 文献中.
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- Cheung WL et al. Acetylation and chromosomal functions. Curr Opin Cell Biol 12:326-33 (2000). PubMed: 10801466
- Strahl BD & Allis CD The language of covalent histone modifications. Nature 403:41-5 (2000). PubMed: 10638745
- Grunstein M Histone acetylation in chromatin structure and transcription. Nature 389:349-52 (1997). PubMed: 9311776