Hello,

I am looking for tyrosinase enzyme inhibitors on B16F10 cells.

Cuurently I am having problems for cellular tyrosinase activity measurement. Which I hav done successfully before.

I dont get any color change on cellular protein extracts however I could see the decrease in the melanin formation.

I measure protein content with Bradford assay. I am using 1% Triton X-100 and 0.1 mM PMSF in pH 6.8 Na-Phosphate buffer.

I am using L-DOPA solution fresh in 5 mM concentration.

My assay is composed of 50 ug/ml protein in 100 ul pH 6.8 tampon and 100 ul 5 mM L-DOPA. I incubate the plates at 37 C for 1 hour.

Is there something I am missing out?Hope my question reaches to some people in the field.