How do you stop the oxidation of Amplex Red? Life Tech sell a proprietary stop reagent but I'd rather use something of which I know the composition. Any ideas? Strong acids will alter fluorescence, alcohols may not be effective. Thanks.
Amplex Red will continue to oxidize over time even after inhibiting HRP, unfortunately. If you have leftover diluted Amplex Red in a tube, at working concentration, put it in a desk drawer and you can see after a few month it turned from colorless to fluorescent pink. If you are looking at fixed H2O2 concentration, your data should be fine as long as you have H2O2 standard curve on the same plate, even if fluorescence numbers are different between runs.
Looking at Molecular Probes' stop reagent data sheet, they only claim to keep fluorescent signal stable for 3 hours. Also the last sentence in the Introduction states that it should work in any other HRP-coupled detection system, so Martin's guess of sodium azide is probably right.
- Badges
- Science topic
- Similar topics
- Chemistry
- Biochemistry