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Recombinant Proteins & Enzymes

Recombinant HDAC6 (H625A) protein

Aliases: HD6, JM21, CPBHM, PPP1R90
Catalog No: 31565 Format: 20 µg $395 Buy Now
Catalog No: 31965 Format: 1 mg $3,500 Buy Now
Expressed In: Baculovirus Protein Species: Human

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for Recombinant HDAC6 (H625A) proteinContents

Recombinant HDAC6 (H625A) 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 (H625A) 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 (H625A) proteinApplication Notes

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

HDAC Activity Assay Conditions:
1 µM H3K9ac (1-21) peptide was incubated with different concentrations of HDAC6 (H625A) protein in a 10 µl reaction system containing 25 mM Tris-HCl pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2 and 0.1 mg/ml BSA for 30 minutes at 37°C. 10 µl H3K9me0 antibody and SA-XL665 mixture (each at 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 minutes. All operations were performed at room temperature. HTRF assay was used for detection.

for Recombinant HDAC6 (H625A) proteinProtein Details

Recombinant human HDAC6 (H625A) was expressed in a baculovirus expression system as the full length protein (accession number NP_001308154.1) with a point mutation His625Ala 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 (H625A) protein gel.

Recombinant HDAC6 (H625A) protein gel
HDAC6 (H625A) protein was run on an 8% SDS-PAGE gel and stained with Coomassie Blue.
MW: 135.7 kDa
Purity: > 90%

HTRF assay for HDAC6 (H625A) activity.

HDAC6 (H625A) Protein activity assay.
1 µM H3K9ac (1-21) peptide was incubated with different concentrations of HDAC6 (H625A) protein in reaction buffer for 30 min. at 37°C followed by developing for 30 min. Reaction products were detected with H3K9me0 antibody. HTRF assay was used for activity detection and shows that H625A mutant loses enzymatic activity.

for Recombinant HDAC6 (H625A) 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 (H625A) 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 (H625A) protein
 

Data Thumbnails

Recombinant HDAC6 (H625A) protein gel.

Recombinant HDAC6 (H625A) protein gel.
(Click to enlarge and see details).

HTRF assay for HDAC6 (H625A) activity.

HTRF assay for HDAC6 (H625A) activity
(Click to enlarge and see details).