Recombinant SIRT3 (102-399) protein

Aliases: SIRT-3, Sirtuin 3, Sir1-Like 3, EC 3.5.1
Catalog No: 31529 Format: 100 µg $410 Buy
Expressed In: E. coli Protein Species: Human

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Contents

Recombinant SIRT3 (102-399) protein is supplied in 25 mM Tris pH 8.0, 150 mM NaCl, 5% glycerol. Please refer to product insert upon arrival for lot-specific concentration. Please refer to product insert upon arrival for lot-specific concentration.

Background

SIRT3 (Sirtuin 3) is a member of the SIR2 / Sirtuin family of NAD+-dependant deacetylase enzymes. Sirtuin family enzymes can be found in the cytoplasm, mitochondria or nucleus of a cell and are ubiquitously expressed. SIRT3 has multiple targets for deacetylation, including ACSS1, IDH, GDH, SOD2, PDHA1, LCAD, SDHA and the ATP synthase subunit ATP5O. SIRT3 contributes to the regulation of the cellular energy metabolism and tissue-specifc ATP levels.

Application Notes

Recombinant SIRT3 protein is useful for histone and other protein deacetylase assays and the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

HDAC Activity Assay Conditions: 3 µM histone peptide H3K9ac (1-21aa) was incubated with SIRT3 (102-399) in reaction buffer including 25 mM Tris-HCl pH 8.0, 137 mM NaCl, 2.7 mM MgCl2, 1 mM KCl, 0.1 mg/ml BSA and 50 µM NAD+ 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 SIRTs - click here to view the list.

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

Protein Details

Recombinant SIRT3 (102-399) protein that includes amino acids 102 - 399 of human SIRT3 protein (accession number NM_012239.5) was expressed in E. coli and contains an N-terminal GST-Tag with a molecular weight of 59.7 kDa. The purity of the protein is > 60% by SDS-PAGE.

 
SDS-PAGE Recombinant SIRT3 (102-399)

Recombinant SIRT3 on 12.5% SDS-PAGE with Coomassie Blue staining.

MW: 64.3 kDa. Purity: > 60%

HTRF assay for SIRT3 (102-399) activity

HTRF assay for SIRT3 (102-399) activity.

3 µM histone peptide H3K9ac (1-21) was incubated with SIRT3 (102-399) in reaction buffer for 30 min at 37°C. Anti-H3K9me0 antibody was used to detect reaction products.

HTRF assay for SIRT3 (102-399) activity

HTRF assay for SIRT3 (102-399) activity.
3 µM histone peptide H3K9ac (1-21) was incubated with SIRT3 (102-399) in reaction buffer for 30 min at 37°C. Anti-H3K9me0 antibody was used to detect reaction products.

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.