Protein carbonyl determination by a rhodamine B hydrazide-based fluorometric assay

Add time:07/25/2019    Source:sciencedirect.com

A new fluorometric assay is presented for the ultrasensitive quantification of total protein carbonyls, and is based on their specific reaction with rhodamine B hydrazide (RBH), and the production of a protein carbonyl-RBH hydrazone the fluorescence of which (at ex/em 560/585 nm) is greatly enhanced by guanidine-HCl. Compared to the fluorescein-5-thiosemicarbazide (FTC)-based fluorometric assay, the RBH assay uses a 24-fold shorter reaction incubation time (1 h) and at least 1000-fold lower protein quantity (2.5 µg), and produces very reliable data that were verified by extensive standardization experiments. The protein carbonyl group detection sensitivity limit of the RBH assay, based on its standard curve, can be as low as 0.4 pmol, and even lower. Counting the very low protein limit of the RBH assay, its cumulative and functional sensitivity is 8500- and 800-fold higher than the corresponding ones for the FTC assay. Neither heme proteins hemoglobin and cytochrome c nor DNA interfere with the RBH assay.

We also recommend Trading Suppliers and Manufacturers of fluorescein hydrazide (cas 109653-47-6). Pls Click Website Link as below: cas 109653-47-6 suppliers

Prev:Selective and sensitive fluorometric determinations of cobalt(II) and hydrogen peroxide with fluorescein-hydrazide
Next:Neocryptotanshinone (cas 109664-02-0) inhibits lipopolysaccharide-induced inflammation in RAW264.7 macrophages by suppression of NF-κB and iNOS signaling pathways)