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HTRF SIRT1 Detection Kit, 1,000 Assay Points

The HTRF SIRT1 assay enables a straigthforward screening and characterization of compounds activating or inhibiting SIRT1 activity

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Feature	Specification
Application	Biochemical Enzymatic Assay
Sample Volume	10 µL

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The HTRF SIRT1 assay enables a straigthforward screening and characterization of compounds activating or inhibiting SIRT1 activity

View product information

Product Variants

Part #:

64SI1PEB

List Price

USD 1,372.23

Your online price:

Part #:

64SI1PEC

List Price

USD 11,542.78

Your online price:

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).

Product information

Overview

SIRT1 belongs to class III of the silent information regulator 2 (Sir2) family of proteins, which are histone/protein deacetylases (sirtuins). The use of SIRT1 modulators has been described as a potential therapeutic solution for treating multiple disease states including cancer, diabetes, aging, and neurodegenerative, metabolic or cardiovascular diseases. This rapid, simple assay enables the discrimination of polyphenolic compounds and can be used for high throughput screening to identify both inhibitors and activators of SIRT1 activity.

Specifications

Other Specifications

Application	Biochemical Enzymatic Assay
Brand	HTRF
Detection Modality	HTRF
Product Group	Kit
Sample Volume	10 µL
Shipping Conditions	Shipped in Dry Ice
Target Class	Epigenetics
Technology	TR-FRET
Therapeutic Area	Metabolism/Diabetes Neuroscience Oncology & Inflammation
Unit Size	1,000 assay points

Video gallery

How it works

Assay principle

The HTRF SIRT1 assay uses an anti-acetyl specific MAb labeled with Eu-cryptate and an acetylated-d2 labeled substrate. The maximum signal is obtained in absence of SIRT1 and decreases proportionally to the deacetylation process. The quantity of deacetylated peptides is then directly correlated to the signal extinction.

Assay protocol

The assay consists of two steps: an enzymatic step followed by a universal detection step. The SIRT1 enzyme is first incubated in the presence or absence of compounds with the acetylated d2 labeled substrate. NAD+ is added to start the reaction. The Eu-cryptate labeled MAb is then added. The detection buffer contains EDTA and a SIRT inhibitor (nicotinamide) to stop the enzymatic detection.

Analytical performance

Screening robustness

Z' values for different SIRT1 concentrations in twenty-four replicates were run using 4 selected SIRT1 concentrations: 0 mU of enzyme (no deacetylation), 100 mU of enzyme (100% deacetylation), 10 mU and 20 mU corresponding to the EC50 and EC80 respectively.

Finding activator and inhibitor compounds

Titration curves were performed using 3 classes of compounds: two polyphenolic compounds (resveratrol and piceatannol), two non-polyphenolic compounds (activator1 and activator2 and one reference inhibitor (nicotinamide).