Description
Key facts
Detection method:Fluorescent,
Sample types:Cell culture extracts, Tissue Extracts,
Reacts with:Mouse, Rat, Human,
Assay type:Enzyme activity,
Assay time:1h,
Assay Platform:Microplate reader
Product details:
SIRT1 Activity Assay Kit (Fluorometric) (ab156065) is used to measure SIRT1 activity in either samples enriched for SIRT1, or in purified SIRT1 protein.
How the assay works
Primarily, the SIRT1 Activity Assay Kit is designed for the rapid and sensitive evaluation of SIRT1 inhibitors or activators using crude SIRT1 fractions or purified SIRT1. Additionally, any cultured primary cell, cell line, or tissue homogenate can be assayed for SIRT1 activity with the SIRT1 Activity Assay Kit, if an anti-SIRT1 antibody is used for immunoprecipitation.
SIRT1 Activity Assay Kit (Fluorometric) ab156065 has been shown to detect the activity of Sirtuins, at least SIRT1 in Human or animal cell lysates or in column fractions. The assay shows good linearity of sample response. The assay may be used to follow the purification of Sirtuins or may be used to detect the presence of Sirtuins in cell lysates.
Applications for this kit include:
1. Monitoring the purification of SIRT1.
2. Screening inhibitors or activators of SIRT1.
3. Detecting the effects of pharmacological agents on SIRT1.
SIRT1 Activity Assay Kit (Fluorometric) protocol summary:
- Prepare samples
- Add inhibitor and developer to appropriate wells
- Add SIRT1 enzyme
- Add Stop Solution to stop reaction
- Measure Fluorescence Intensity at 1 – 2 min intervals at Ex/ Em = 350/ 460 nm
To quantify total SIRT enzyme activity, we recommend Universal SIRT Activity Assay Kit (Colorimetric)
ab156915
How other researchers are using ab156065
SIRT1 Activity Assay Kit (Fluorometric) has been used with a variety of sample types including:
- Mouse Liver cells 1
- Human skin fibroblasts 2
- Human Hepatocellular carcinoma cells 3
References: 1 - Yang J et al. 2023; 2 - Thakur B et al. 2023; 3 - Guo S et al. 2023.
Properties and Storage Information:
Shipped at conditions-Dry Ice, Appropriate short-term storage conditions--80°C, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
SIRT1 also known as Silent mating type information regulation 2 homolog 1 is a NAD-dependent deacetylase enzyme. SIRT1 weighs approximately 120 kDa and plays an important role in regulating transcription apoptosis and stress resistance. Researchers have found SIRT1 in various tissues with higher expression in the heart brain and skeletal muscle. It is a component of the larger family of sirtuins which are involved in metabolic regulation and aging.
Biological function summary
SIRT1 modulates several cellular processes such as gene silencing DNA repair and lifespan extension. SIRT1 participates in complexes with other proteins including histones and transcription factors to influence chromatin structure and gene expression. It acts through deacetylation of target proteins affecting their function and stability. The activity of SIRT1 is also linked to environmental and cellular conditions including caloric intake and oxidative stress.
Pathways
SIRT1 is integral in the regulation of metabolic and longevity pathways. It interacts with the FOXO family proteins and the tumor suppressor protein p53 aiding in response to cellular stress and metabolic demands. The role of SIRT1 in the insulin signaling pathway exemplifies its influence on glucose homeostasis and energy balance. These interactions highlight its importance in metabolic health and aging.
SIRT1 links to neurodegenerative diseases such as Alzheimer's disease and metabolic disorders like type 2 diabetes. In Alzheimer's disease SIRT1 interacts with the amyloid precursor protein suggesting a protective role against amyloid-beta accumulation. Additionally studies have shown connections between SIRT1 and insulin receptor substrates highlighting its role in managing insulin sensitivity and glucose metabolism in diabetes. Understanding SIRT1's functions offers potential therapeutic targets for these disorders.
