Recombinant HDAC8 protein, His-Tag

Aliases: Histone Deacetylase 8, HDACL1, HD8, CDA07, CDLS5, MRXS6, RPD3
Catalog No: 31566 Format: 50 µg $450 Buy
Catalog No: 31966 Format: 1 mg $4,050 Buy

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Expressed In: Baculovirus Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

HDAC8 (Histone Deacetylase 8) is a member of the class I mammalian histone deacetylases (HDACs) involved in regulating chromatin structure during transcription. These enzymes catalyze the removal of acetyl groups from lysine residues of histones and other cellular proteins. Lysine N-ε-acetylation is a dynamic, reversible and tightly regulated protein and histone modification that plays a major role in regulation of gene expression in various cellular functions. It consists of the transfer of an acetyl moiety from an acetyl coenzyme A to the ε-amino group of a lysine residue. In vivo, acetylation is controlled by the antagonistic activities of histone acetyltransferases (HATs) and histone deacetylases (HDACs). The HDACs are grouped into four classes, on the basis of similarity to yeast counterparts: HDAC class I (HDAC1, HDAC2, HDAC3 and HDAC8), class II (HDAC4, HDAC5, HDAC6, HDAC7, 9 and 10), class III (SIRT1-7) and class IV (HDAC11). HDAC8 catalyzes the deacetylation of lysine residues in the histone N-terminal tails and represses transcription in large multiprotein complexes with transcriptional co-repressors. It also is involved in the deacetylation of cohesin complex protein SMC3 regulating release of cohesin complexes from chromatin. HDAC8 may play a role in smooth muscle cell contractility.

Application Notes

This protein is useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

HDAC Activity Assay Conditions:
3 µM Histone H3K9ac (1-21 aa) peptide was incubated with HDAC8 protein in reaction buffer including 25 mM Tris-HCl pH 8.0, 137 mM NaCl, 2.7 mM MgCl2, 1 mM KCl and 0.1 mg/ml BSA for 30 min at 37°C. HTRF assay was used for activity detection.

Active Motif has a collection of small-molecule epigenetic modulators that target HDACs - click here to view the list.

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Protein Details

Recombinant human HDAC8 protein, His-Tag was expressed in a baculovirus expression system as the full length protein (accession number NP_060956.1) with a C-terminal 6xHis Tag. The molecular weight of the protein is 43.8 kDa. The HDAC8 protein, His-Tag is >90% pure by SDS-PAGE.

 

Recombinant HDAC8 His-Tag protein gel. HDAC8 protein was run on an 8% SDS-PAGE gel and stained with Coomassie Blue.

HTRF assay for HDAC8 protein, His-Tag
3 µM Histone H3K9ac (1-21aa) peptide was incubated with HDAC8 protein, His-Tag in reaction buffer for 30 min at 37°C, Reaction product was detected by Anti-H3K9me0-Eu antibody.
HTRF assay was used for activity detection.

HTRF assay for HDAC8 protein, His-Tag
3 µM Histone H3K9ac (1-21aa) peptide was incubated with HDAC8 protein, His-Tag in reaction buffer for 30 min at 37°C, Reaction product was detected by Anti-H3K9me0-Eu antibody.
HTRF assay was used for activity detection.

Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.