melanin assay and tyrosinase activity

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Meedo's picture
melanin assay and tyrosinase activity

I'm assying the ability of melanocytes to produce melanin in response to different drugs but with the protocol I use, I usually find high basal level of melanin content in comparison with the stimulating drugs. Can you help me to find the cause of such high basal level and how can I overcome this problem. Another thing, Can you please help me find a clear protocol detecting tyrosinase activity using radioactive method for example the use of triteted tyrosine in order to look for the tritiated H2O. I tried this method but I usually find no response to my stimulating drugs although they are expected to increase the enzyme activity (ang again the basal level is usually high).
N.B I used a postive control that is evident to increase meanin content and tyrosinase activity but I got the same results.
please needs your help ASAP.