For this you have to optimise it’s concentration not someone may tell you for laccase assays . You may start with 0.2 mM with various concentration

& optimise it better to get accurate results

Thanks a lot!

Dear Nasim,

I don't see the necessity to use H2O2 for laccase! Working in aerobic condition could be enough for the ABTS assay.

Regarding the laccase, first of all you should determinate the protein concentration in the filtrate : if it's concentrated you can begin by 10-20 µl , if not you should use huge quantity such 50µl but don't exceed 100 µl.

For ABTS you can using 0.1 or 0.2 mM of ABTS to obtain good dDO for you assays. High concentration of ABTS could abberate the initial OD and can be followed by very quick oxidation !

For the acetate buffer I using alawys 50mM solution, the pH 5.0 is the best, good choice.

Good luck :)

Thanks a lot! :)