Histone Deacetylase Assay Kit (HDAC)

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.

Kit capacity: 100 assays

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

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%.

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