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
Product nameSIRT3 Activity Assay Kit (Fluorometric)
Sample typeCell culture extracts, Tissue Extracts
Assay typeEnzyme activity
Assay time0h 40m
Species reactivityReacts with: Human
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
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.
Storage instructionsPlease 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
- Epigenetics and Nuclear Signaling
- Chromatin Modifying Enzymes
- Class III / Sir2 class
FunctionNAD-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 specificityWidely expressed.
Sequence similaritiesBelongs to the sirtuin family. Class I subfamily.
Contains 1 deacetylase sirtuin-type domain.
modificationsProcessed by mitochondrial processing peptidase (MPP) to give a 28 kDa product. Such processing is probably essential for its enzymatic activity.
Cellular localizationMitochondrion matrix.
- Information by UniProt
- hSIRT 3
- Mitochondrial nicotinamide adenine dinucleotide dependent deacetylase
ab156067 has been referenced in 8 publications.
- Botezelli JD et al. Adipose depot-specific upregulation of Ucp1 or mitochondrial oxidative complex proteins are early consequences of genetic insulin reduction in mice. Am J Physiol Endocrinol Metab 319:E529-E539 (2020). PubMed: 32715748
- Ouyang J et al. SIRT3 Inactivation Promotes Acute Kidney Injury Through Elevated Acetylation of SOD2 and p53. J Surg Res 233:221-230 (2019). PubMed: 30502252
- Miao HH et al. Ginsenoside Rg1 attenuates isoflurane/surgery-induced cognitive disorders and sirtuin 3 dysfunction. Biosci Rep 39:N/A (2019). PubMed: 31652451
- Yi X et al. SIRT3-Dependent Mitochondrial Dynamics Remodeling Contributes to Oxidative Stress-Induced Melanocyte Degeneration in Vitiligo. Theranostics 9:1614-1633 (2019). PubMed: 31037127
- Chen LY et al. Activation of AMPK-SIRT3 signaling is chondroprotective by preserving mitochondrial DNA integrity and function. Osteoarthritis Cartilage 26:1539-1550 (2018). PubMed: 30031925
- Yin J et al. Sirtuin 3 attenuates amyloid-ß induced neuronal hypometabolism. Aging (Albany NY) 10:2874-2883 (2018). PubMed: 30362958
- Michalak S et al. Mitochondrial Respiration in Intact Peripheral Blood Mononuclear Cells and Sirtuin 3 Activity in Patients with Movement Disorders. Oxid Med Cell Longev 2017:9703574 (2017). PubMed: 29081897
- Kwon S et al. Obesity and aging diminish sirtuin 1 (SIRT1)-mediated deacetylation of SIRT3, leading to hyperacetylation and decreased activity and stability of SIRT3. J Biol Chem 292:17312-17323 (2017). PubMed: 28808064