Revvity, 64SI1PEB, HTRF SIRT1 Detection Kit, 1,000 Assay Points

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.
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
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. 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