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SIRT3 Activity Assay Kit (Fluorometric) (ab156067) detects SIRT3 activity in lysates.

Is this your first time purchasing this assay kit? Get 20% off one kit by entering code 20OFF-T5A8Q at checkout. T&C’s apply – click here. Offer only valid when purchasing directly from Abcam in Europe, North America, Australia, and Singapore.


Images

Key facts

Detection method

Fluorescent

Sample types

Cell culture extracts, Tissue Extracts

Assay type

Enzyme activity

Reactive species

Human

Assay time

40m

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Target data

Function

NAD-dependent protein deacetylase (PubMed:12186850, PubMed:12374852, PubMed:16788062, PubMed:18680753, PubMed:18794531, PubMed:23283301, PubMed:24121500, PubMed:24252090, PubMed:19535340). Activates or deactivates mitochondrial target proteins by deacetylating key lysine residues (PubMed:12186850, PubMed:12374852, PubMed:16788062, PubMed:18680753, PubMed:18794531, PubMed:23283301, PubMed:24121500, PubMed:24252090). Known targets include ACSS1, IDH, GDH, SOD2, PDHA1, LCAD, SDHA and the ATP synthase subunit ATP5PO (PubMed:16788062, PubMed:18680753, PubMed:24121500, PubMed:24252090, PubMed:19535340). Contributes to the regulation of the cellular energy metabolism (PubMed:24252090). Important for regulating tissue-specific ATP levels (PubMed:18794531). In response to metabolic stress, deacetylates transcription factor FOXO3 and recruits FOXO3 and mitochondrial RNA polymerase POLRMT to mtDNA to promote mtDNA transcription (PubMed:23283301). Acts as a regulator of ceramide metabolism by mediating deacetylation of ceramide synthases CERS1, CERS2 and CERS6, thereby increasing their activity and promoting mitochondrial ceramide accumulation (By similarity).

Alternative names

What's included?

100 Test
Components
Developer
1 x 500 µL
Fluoro-Deacetylated Peptide (0.2 mM)
1 x 100 µL
Fluoro-Substrate Peptide (0.2 mM)
1 x 500 µL
NAD (2 mM)
1 x 500 µL
Recombinant SIRT3
1 x 500 µL
SIRT Assay Buffer
2 x 1 mL
Stop Solution
2 x 1 mL

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SIRT3 Activity Assay Kit (Fluorometric) (ab156067) detects SIRT3 activity in lysates.

Is this your first time purchasing this assay kit? Get 20% off one kit by entering code 20OFF-T5A8Q at checkout. T&C’s apply – click here. Offer only valid when purchasing directly from Abcam in Europe, North America, Australia, and Singapore.

Alternative names

Key facts

Detection method

Fluorescent

Sample types

Cell culture extracts, Tissue Extracts

Assay type

Enzyme activity

Reactive species

Human

Assay time

40m

Assay Platform

Microplate reader

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-80°C

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

Notes

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.

Supplementary info

Biological function summary

SIRT3 modulates energy homeostasis by deacetylating and regulating enzymes involved in metabolism. It is a part of the mitochondrial sirtuin family contributing to fatty acid oxidation and the antioxidant defense system. By targeting key metabolic enzymes SIRT3 directly impacts the tricarboxylic acid (TCA) cycle and oxidative phosphorylation processes. These functions highlight its role in maintaining energy efficiency and reducing reactive oxidative species.

Activity summary

SIRT3 also known as Sirtuin 3 and mitochondrial sirtuin is a deacetylase with a mass of roughly 44 kDa. This protein is mostly found in the mitochondria where it regulates the acetylation state of various substrates. It plays an important role in maintaining mitochondrial homeostasis by modifying protein activity. SIRT3 is evolutionarily conserved and highly expressed in tissues with high energy demand like the heart liver and brown adipose tissue.

Pathways

SIRT3 participates in vital metabolic pathways such as the TCA cycle and the urea cycle. It interacts with other proteins like MnSOD and IDH2 to enhance mitochondrial function and overall cellular metabolism. This helps control the balance between energy production and consumption integrating signals from the AMPK and NAD+-dependent pathways to coordinate responses to changes in energy availability.

Associated diseases and disorders

SIRT3 has implications in metabolic syndrome and cancer. Reduced SIRT3 activity is associated with increased susceptibility to metabolic disturbances like obesity and type 2 diabetes. Its deficiency affects the regulation of ROS and metabolic adaptation linking it to tumorigenesis and cancer progression. As it connects with proteins like HIF-1α in hypoxic conditions SIRT3 plays a role in cellular adaptation to stress influencing disease progression.

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2 product images

  • Functional Studies - SIRT3 Activity Assay Kit (Fluorometric) (ab156067), expandable thumbnail

    Functional Studies - SIRT3 Activity Assay Kit (Fluorometric) (ab156067)

    Dose dependency curve of recombinant SIRT3 activity

  • Functional Studies - SIRT3 Activity Assay Kit (Fluorometric) (ab156067), expandable thumbnail

    Functional Studies - SIRT3 Activity Assay Kit (Fluorometric) (ab156067)

    Effect of Trichostatin A and NAD on recombinant SIRT3 activity

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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