SIRT3 Activity Assay Kit (Fluorometric) (ab156067)
Key features and details
- Assay type: Enzyme activity
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 40 min
- Sample type: Cell culture extracts, Tissue Extracts
Overview
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Product name
SIRT3 Activity Assay Kit (Fluorometric) -
Detection method
Fluorescent -
Sample type
Cell culture extracts, Tissue Extracts -
Assay type
Enzyme activity -
Assay time
0h 40m -
Species reactivity
Reacts with: Human -
Product overview
Abcam’s SIRT3 Activity Assay Kit (Fluorometric) (ab156067) detects SIRT3 activity in lysates.
Primarily, the SIRT3 Activity Assay Kit (Fluorometric) is designed for the rapid and sensitive evaluation of SIRT3 inhibitors or activators using crude SIRT3 fraction or purified SIRT3.
Applications for this kit include:
1. Screening inhibitors or activators of SIRT3.
2. Detecting the effects of pharmacological agents on SIRT3.
SIRT3 assay protocol summary:
- add assay buffer, substrate peptide and NAD to wells
- add developer to wells
- add enzyme sample or recombinant SIRT3 to wells
- analyze with microplate reader for 30-60 min every 1-2 min -
Notes
Histone Deacetylases (HDACs) are a class of enzymes responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4), allowing the histones to wrap the DNA more tightly.
HDAC proteins occur in four groups (class I, class IIA, class IIB, class III, class IV) based on function and DNA sequence similarity.
Classes I, IIA and IIB are considered "classical" HDACs whose activities are inhibited by trichostatin A (TSA), whereas class III is a family of NAD+-dependent proteins (sirtuins) not affected by TSA. Class IV is considered an atypical class on its own, based solely on DNA sequence similarity to the others. -
Platform
Microplate reader
Properties
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Storage instructions
Please refer to protocols. -
Components 100 tests Developer 1 x 500µl Fluoro-Deacetylated Peptide (0.2 mM) 1 x 100µl Fluoro-Substrate Peptide (0.2 mM) 1 x 500µl NAD (2 mM) 1 x 500µl Recombinant SIRT3 1 x 500µl SIRT Assay Buffer 2 x 1ml Stop Solution 2 x 1ml -
Research areas
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Function
NAD-dependent protein deacetylase. Activates or deactivates mitochondrial target proteins by deacetylating key lysine residues. Known targets include ACSS1, IDH, GDH, SOD2, PDHA1, LCAD, SDHA and the ATP synthase subunit ATP5O. Contributes to the regulation of the cellular energy metabolism. Important for regulating tissue-specific ATP levels. -
Tissue specificity
Widely expressed. -
Sequence similarities
Belongs to the sirtuin family. Class I subfamily.
Contains 1 deacetylase sirtuin-type domain. -
Post-translational
modificationsProcessed by mitochondrial processing peptidase (MPP) to give a 28 kDa product. Such processing is probably essential for its enzymatic activity. -
Cellular localization
Mitochondrion matrix. - Information by UniProt
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Alternative names
- hSIRT 3
- hSIRT3
- Mitochondrial nicotinamide adenine dinucleotide dependent deacetylase
see all -
Database links
- Entrez Gene: 23410 Human
- Omim: 604481 Human
- SwissProt: Q9NTG7 Human
- Unigene: 716456 Human
Images
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ab156067 - SIRT3 Activity Assay Kit (Fluorometric)Dose dependency curve of recombinant SIRT3 activity
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ab156067 - SIRT3 Activity Assay Kit (Fluorometric)Effect of Trichostatin A and NAD on recombinant SIRT3 activity