What is the best way to quantitate Cyclic ADP-Ribose (cADPR)?
Posted June 13, 2019
I am using a fluorescence microplate reader.
We recommend using our Amplite® Fluorimetric cADP-Ribose Assay Kit (Cat# 20305) to quantitate cADPR. This assay is convenient for either 96-well or 384-well microtiter plate format. In tissues and cell cultures, it can detect cADPR in the low nM range. The lowest detected concentration of cADPR is 100 nM.
How Cat#20305 works:
cADPR is derived from NAD+ via ADP-riboxyl cyclase catalyzation. This reaction can be reversed in the presence of high concentration of nicotinamide, producing NAD+ from cADPR stoichiometrically. The resulting NAD+ is measured with NAD sensor, Quest Fluor™ NAD Probe. This probe is highly sensitive for NAD+ and has no reaction to NADH. The fluorescent signal can be readily detected at Ex/Em = 420/480 nm.
Amplite™ Fluorimetric cADP-Ribose Assay product page