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Histone Deacetylase Assay Kit (HDAC)
Description:
Histone Deacetylase Assay Kit (HDAC)
Trade Name:
Upstate (Millipore)
Qty/Pk:
1 kit
Product Overview:
This kit contains nonradioactive components for radiolabeling and purifying histone H4 peptide with [3H]-acetyl CoA. The radiolabeled peptide is a suitable substrate for assay of HDAC activity. HeLa nuclear extract, which is rich in histone deacetylase activity, is included as a positive control. The HDAC inhibitor sodium butyrate is provided to demonstrate specificity of the deacetylase activity.
Key Applications:
Enzyme Assay
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Entrez Gene Summary:
Histones play a critical role in transcriptional regulation, cell cycle progression, and developmental events. Histone acetylation/deacetylation alters chromosome structure and affects transcription factor access to DNA. The protein encoded by this gene belongs to the class II histone deacetylase/acuc/apha family. It possesses histone deacetylase activity and represses transcription when tethered to a promoter. It coimmunoprecipitates only with HDAC3 family member and might form multicomplex proteins. It also interacts with myocyte enhancer factor-2 (MEF2) proteins, resulting in repression of MEF2-dependent genes. This gene is thought to be associated with colon cancer. Two transcript variants encoding different isoforms have been found for this gene.
UniProt Summary:
FUNCTION: SwissProt: Q9UQL6 # 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. Involved in muscle maturation by repressing transcription of myocyte enhancer MEF2C. During muscle differentiation, it shuttles into the cytoplasm, allowing the expression of myocyte enhancer factors.
SIZE: 1122 amino acids; 121992 Da
SUBUNIT: Interacts with BCOR, HDAC7, HDAC9, CTBP1, MEF2C, NCOR2, NRIP1, PHB2 and a 14-3-3 chaperone protein. Interacts with JARID1B.
SUBCELLULAR LOCATION: Nucleus. Cytoplasm. Note=Shuttles between the nucleus and the cytoplasm. In muscle cells, it shuttles into the cytoplasm during myocyte differentiation. The export to cytoplasm depends on the interaction with a 14-3-3 chaperone protein and is due to its phosphorylation at Ser-259 and Ser-498 by CaMK.TISSUE SPECIFICITY: Ubiquitous.DOMAIN:SwissProt: Q9UQL6 The nuclear export sequence mediates the shuttling between the nucleus and the cytoplasm (By similarity).
PTM: Phosphorylated by CaMK at Ser-259 and Ser-498. The phosphorylation is required for the export to the cytoplasm. & Ubiquitinated. Polyubiquitination however does not lead to its degradation.
SIMILARITY: Belongs to the histone deacetylase family. Type 2 subfamily.
SIZE: 1122 amino acids; 121992 Da
SUBUNIT: Interacts with BCOR, HDAC7, HDAC9, CTBP1, MEF2C, NCOR2, NRIP1, PHB2 and a 14-3-3 chaperone protein. Interacts with JARID1B.
SUBCELLULAR LOCATION: Nucleus. Cytoplasm. Note=Shuttles between the nucleus and the cytoplasm. In muscle cells, it shuttles into the cytoplasm during myocyte differentiation. The export to cytoplasm depends on the interaction with a 14-3-3 chaperone protein and is due to its phosphorylation at Ser-259 and Ser-498 by CaMK.TISSUE SPECIFICITY: Ubiquitous.DOMAIN:SwissProt: Q9UQL6 The nuclear export sequence mediates the shuttling between the nucleus and the cytoplasm (By similarity).
PTM: Phosphorylated by CaMK at Ser-259 and Ser-498. The phosphorylation is required for the export to the cytoplasm. & Ubiquitinated. Polyubiquitination however does not lead to its degradation.
SIMILARITY: Belongs to the histone deacetylase family. Type 2 subfamily.
Quality Assurance:
Routinely evaluated with two experiments, The dependence of HDAC activity on HeLa Nuclear Extract and the inhibition of HDAC Activity by Sodium Butyrate.
In the first experiment, HeLa nuclear extract was incubated at room temperature with [3H]-acetyl-Histone H4 peptide. Released [3H]-acetate was measured using a scintillation counter.
In the second experiment, [3H]-acetyl-Histone H4 peptide was incubated with HeLa nuclear extract in the presence or absence of Sodium Butyrate. Sodium Butyrate inhibited release of [3H]-acetate by ~92%.
In the first experiment, HeLa nuclear extract was incubated at room temperature with [3H]-acetyl-Histone H4 peptide. Released [3H]-acetate was measured using a scintillation counter.
In the second experiment, [3H]-acetyl-Histone H4 peptide was incubated with HeLa nuclear extract in the presence or absence of Sodium Butyrate. Sodium Butyrate inhibited release of [3H]-acetate by ~92%.
Components:
- HeLa Nuclear Extract (Cat.# 12-309)
- Histone H4 Peptide, biotin conjugate, residues 2-24 (Cat.# 12-372)
- HAT Assay Buffer, 5X (Cat.# 20-148)
- Sodium Butyrate (Cat.# 19-137)
- 5X HDAC Assay Buffer
- Streptavidin Agarose
- PCAF active
Brand Family:
Upstate
UniProt Number:
Packaging:
Kit capacity: 100 assays
Protein/Isoform Description:
Histone Deacetylases reverse the acetylation of lysines that are effected by HATs. Class I, II and III HDACs are homologous to yeast proteins Rpd3, Hda1 and Sir2, respectively. HDAC activity represses gene expression, and inhibition of HDACs can sometimes reactivate silenced genes. HDACs represent an important new class of drug targets.
Gene Symbol:
- HDAC1
- RPD3L1
- GON-10
- DKFZp686H12203
- HD1
- RPD3
Product Name:
Histone Deacetylase Assay Kit (HDAC)
Entrez Gene Number: