Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate (12 x 8 well strips)
产品名称HDAC Activity Assay试剂盒(colorimetric)
参阅全部 HDAC 试剂盒
实验步骤Multiple steps standard assay
Inhibition of histone deacetylases (HDACs) has been implicated in the modulation of transcription and the induction of apoptosis or differentiation in cancer cells. However, screening HDAC inhibitory compounds has proven to be difficult over the past due to the lack of convenient tools for analyzing HDAC activity. The new Colorimetric HDAC Activity Assay Kit ab1432 provides a fast and convenient colorimetric method that eliminates radioactivity, extractions, or chromatography, as used in the traditional assays. The new method requires only two easy steps, both performed on the same microtiter plate. First, the HDAC colorimetric substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (e.g., HeLa nuclear extract or your own samples). Deacetylation of the substrate sensitizes the substrate, so that, in the second step, treatment with the Lysine Developer produces a chromophore. The chromophore can be easily analyzed using an ELISA plate reader or spectrophotometer. The assay is well suited for high throughput screening applications.
This product is manufactured by BioVision, an Abcam company and was previously called K331 HDAC Activity Colorimetric Assay Kit. K331-100 is the same size as the 96 test size of ab1432.
Read the entire protocol (attached to the datasheet) before beginning the procedure.
The HeLa extract should be refrozen immediately at -20 or -70°C after each use to avoid loss of activity. The Lysine Developer should be refrozen immediately at –20 or -70°C after each use or aliquotted for future use.
This kit contains enough reagents for about 100 tests. If positive and negative controls are included, 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.
经测试应用适用于: Functional Studiesmore details
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 (10 mM) Yellow Cap 1 x 20µl HDAC Inhibitor (Trichostatin A, 1 mM) Blue Cap 1 x 10µl HDAC Substrate Amber Cap 1 x 500µl HeLa Nuclear Extract (5 mg/mL) Red Cap 1 x 50µl 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
The Abpromise guarantee
Use at an assay dependent concentration.
Use at an assay dependent concentration.
ab1432 被引用在 13 文献中.
- Kuroda N et al. Apoptotic response through a high mobility box 1 protein-dependent mechanism in LPS/GalN-induced mouse liver failure and glycyrrhizin-mediated inhibition. PLoS One 9:e92884 (2014). PubMed: 24690901
- Evankovich J et al. High Mobility Group Box 1 Release from Hepatocytes during Ischemia and Reperfusion Injury Is Mediated by Decreased Histone Deacetylase Activity. J Biol Chem 285:39888-97 (2010). PubMed: 20937823
- Wu Y et al. IIp45 Inhibits Cell Migration through Inhibition of HDAC6. J Biol Chem 285:3554-60 (2010). PubMed: 20008322
- Imesch P et al. Romidepsin reduces histone deacetylase activity, induces acetylation of histones, inhibits proliferation, and activates apoptosis in immortalized epithelial endometriotic cells. Fertil Steril 94:2838-42 (2010). PubMed: 20605144
- Dedes KJ et al. Acquired vorinostat resistance shows partial cross-resistance to 'second-generation' HDAC inhibitors and correlates with loss of histone acetylation and apoptosis but not with altered HDAC and HAT activities. Anticancer Drugs 20:321-33 (2009). PubMed: 19322073