This plan only provides a guide, please modify it to meet your specific needs.
PBD-BODIPY is a probe for spectrophotometric measurements of autoxidation reactions.
Application case: PBD-BODIPY as signal carrier in styrene autoxidation
Literature source:
Omkar Zilka, et al.On the Mechanism of Cytoprotection by Ferrostatin-1 and Liproxstatin-1 and the Role of Lipid Peroxidation in Ferroptotic Cell Death. ACS Cent Sci. 2017 Mar 22;3(3):232-243. doi: 10.1021 /acscentsci.7b00028. Epub 2017 Mar 7.
experimental method:
1. Styrene was washed three times with 1 M aqueous NaOH, dried over MgSO4, filtered, vacuum distilled, and purified by precipitation of silica and then basic alumina;
2. Add 1.18 mL of chlorobenzene to the cuvette containing 1.25 mL of styrene, and equilibrate the solution at 37°C for 5 min;
3. To a blank cuvette, add 12.5 μL of 2 mM PBD-BODIPY in 1,2,4-trichlorobenzene, then add 50 μL of 0.3 M AIBN in chlorobenzene, and mix the solution thoroughly;
4.After 20 minutes, add Lip-1, Fer-1, C15-THN, PMHC, or α-TOH in chlorobenzene stock solution (1 mM), followed by loss of absorbance at 591 nm. Co-autoxidation of the PBD-BODIPY signal carrier and hydrocarbon co-substrate can be quantified by detecting the loss of absorbance at 591 nm.
Example of experimental results:
PBD-BODIPY serves as the signal carrier in styrene autoxidations (A), enabling determination of rate constants (kinh) and stoichiometries (n) for reactions of inhibitors with chain-carrying peroxyl radicals (B). Coautoxidations of styrene (4.3M) and PBD-BODIPY (10 μM) initiated by AIBN (6 mM) in chlorobenzene at 37 °C (black trace) and inhibited by 2 μM (red trace), 3 μM (green trace), and 4 μM (blue trace) of Fer-1 (C), Lip-1 (D), C15-THN (E), α-TOH (F), and PMHC (G). Average inhibition rate constants and stoichiometry summarized in panel H. Reaction progress was monitored by absorbance at 591 nm (ε = 139,000 M–1 cm–1). aFrom ref (29); the inhibition rate constant is greater than what can be determined from the data in panel E.
Precautions:
1) Fluorescent dyes all have quenching problems. Please avoid light as much as possible to slow down fluorescence quenching.
2) For your safety and health, please wear a lab coat and disposable gloves.