Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate (12 x 8 well strips)
Product nameHDAC Activity Assay Kit (colorimetric)
See all HDAC kits
Assay durationMultiple steps standard assay
Species reactivityReacts with: Mammals
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.
Tested applicationsSuitable for: Functional Studiesmore details
PlatformMicroplate (12 x 8 well strips)
Storage instructionsStore at -80°C. Please refer to protocols.
Components Identifier 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
FunctionResponsible 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.
Tissue specificityUbiquitous, with higher levels in heart, pancreas and testis, and lower levels in kidney and brain.
Sequence similaritiesBelongs to the histone deacetylase family. HD type 1 subfamily.
modificationsSumoylated 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
Our Abpromise guarantee covers the use of ab1432 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use at an assay dependent concentration.
ab1432 has been referenced in 10 publications.
- 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
- Margariti A et al. Splicing of HDAC7 modulates the SRF-myocardin complex during stem-cell differentiation towards smooth muscle cells. J Cell Sci 122:460-70 (2009). PubMed: 19174469
- Choo MK et al. NFATc1 mediates HDAC-dependent transcriptional repression of osteocalcin expression during osteoblast differentiation. Bone 45:579-89 (2009). PubMed: 19463978
- Imesch P et al. MLH1 protects from resistance acquisition by the histone deacetylase inhibitor trichostatin A in colon tumor cells. Int J Oncol 35:631-40 (2009). PubMed: 19639184
- Chiu J et al. PARP mediates structural alterations in diabetic cardiomyopathy. J Mol Cell Cardiol 45:385-93 (2008). PubMed: 18657544
- Pong RC et al. Mechanism of increased coxsackie and adenovirus receptor gene expression and adenovirus uptake by phytoestrogen and histone deacetylase inhibitor in human bladder cancer cells and the potential clinical application. Cancer Res 66:8822-8 (2006). PubMed: 16951199
- Thangaraju M et al. SLC5A8 triggers tumor cell apoptosis through pyruvate-dependent inhibition of histone deacetylases. Cancer Res 66:11560-4 (2006). PubMed: 17178845
- Zeng L et al. HDAC3 is crucial in shear- and VEGF-induced stem cell differentiation toward endothelial cells. J Cell Biol 174:1059-69 (2006). PubMed: 16982804
- Marwick JA et al. Cigarette smoke alters chromatin remodeling and induces proinflammatory genes in rat lungs. Am J Respir Cell Mol Biol 31:633-42 (2004). PubMed: 15333327