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Recombinant Proteins & Enzymes

Recombinant HDAC6 (H230A) protein

Aliases: HD6, JM21, CPBHM, PPP1R90
Catalog No: 31564 Format: 20 µg $395 Buy Now
Expressed In: Baculovirus Protein Species: Human

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for Recombinant HDAC6 (H230A) proteinContents

Recombinant HDAC6 (H230A) protein is supplied in 25 mM HEPES pH 7.5, 300 mM NaCl, 5% Glycerol, 0.04% Triton X-100, 0.2 mM TCEP. Please refer to product insert upon arrival for lot-specific concentration.

for Recombinant HDAC6 (H230A) proteinBackground

HDAC6 (Histone Deacetylase 6) is a member of the class IIb mammalian histone deacetylases (HDACs) involved in regulating chromatin structure during transcription. These enzymes catalyze the removal of acetyl groups from lysine residues of histones or many cellular proteins. Lysine N-ε-acetylation is a dynamic, reversible and tightly regulated protein and histone modifiation 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). HDAC6 is a unique enzyme that harbors a full duplication of its deacetylase homology region, which appears to contribute independently to the overall activity of HDAC6 protein. alpha-tubulin and HSP90 are two known substrates of HDAC6, playing an important role in cellular mechanisms related to the microtubule network and HSP90-dependent events. HDAC6 also participates in regulating expression of a group of genes involved in the remodeling of chromatin during cell differentiation.

for Recombinant HDAC6 (H230A) proteinApplication Notes

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

HDAC Activity Assay Conditions:
3 µM H3K9ac (1-21) peptides was incubated with different concentrations of HDAC6 (H230A) in 25 mM Tris-HCl, pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 0.1 mg/ml BSA for 30 minutes at 37°C followed by developing for 30 minutes at room temperature.

for Recombinant HDAC6 (H230A) proteinProtein Details

Recombinant human HDAC6 (H230A) was expressed in a baculovirus expression system as the full length protein (accession number NP_001308154.1) with a point mutation His230Ala and a C-terminal FLAG-Tag. The molecular weight of the protein is 135.7 kDa. The recombinant protein is >85% pure by SDS-PAGE.

 
Recombinant HDAC6 (H230A) protein gel.

Recombinant HDAC6 (H230A) protein gel
HDAC6 (H230A) protein was run on an 8% SDS-PAGE gel and stained with Coomassie Blue.

HTRF assay for HDAC6 (H230A) activity.

HDAC6 (H230A) Protein activity assay.
3 µM H3K9ac (1-21) peptides was incubated with different concentrations of HDAC6 (H230A) protein in reaction buffer for 30 minutes at 37°C followed by developing for 30 minutes at room temperature. Reaction product was detected by Anti-H3K9me0-Eu antibody. HTRF assay was used for activity detection.

HTRF assay for HDAC6 (H230A) activity.

HDAC6 (H230A) Protein activity assay.
3 µM H3K9ac (1-21) peptides was incubated with different concentrations of HDAC6 (H230A) protein in reaction buffer for 30 minutes at 37°C followed by developing for 30 minutes at room temperature. Reaction product was detected by Anti-H3K9me0-Eu antibody. HTRF assay was used for activity detection.

for Recombinant HDAC6 (H230A) proteinStorage

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.

for Recombinant HDAC6 (H230A) proteinGuarantee

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.

 

Technical Data Sheet

Recombinant HDAC6 (H230A) protein
 

Data Thumbnails

Recombinant HDAC6 (H230A) protein gel.

Recombinant HDAC6 (H230A) protein gel.
HDAC6 (H230A) protein was run on an 8% SDS-PAGE gel and stained with Coomassie blue.
(Click to enlarge and see details)

HTRF assay for HDAC6 (H230A) activity.

HTRF assay for HDAC6 (H230A) activity
H3K9ac peptides were incubated with HDAC6 (H230A) protein. Reaction product was detected by Anti-H3K9me0-Eu antibody.
(Click to enlarge and see details).

HTRF assay for HDAC6 (H230A) activity.

HTRF assay for HDAC6 (H230A) activity
H3K9ac peptides were incubated with HDAC6 (H230A) protein. Reaction product was detected by Anti-H3K9me0-Eu antibody.
(Click to enlarge and see details).