Research guidance on cell-based whitening assays - (Oct/05/2008 )
I'm a master student and I'm currently trying to develop cell-based whitening assays. However, I'm new to these assays...I read from research papers abt this field of study...It looks simple when read but when I want to work it out, I failed.
Anybody got any experience abt cell-based whitening assays (e.g cellular tyrosinase assay, melanin assay....)? Pls give me some guidance or suggestions.
Could you explain more abt the whitening assay? I have no idea.
Basically I cultured the melanoma cells (B16F10 murine melanoma cells) and used the cells for the assays. Tyrosinase enzyme is the key enzyme important for melanogenesis (melanin formation). So basically need to extract the enzyme from the cells, do protein quantification, then carry out enzyme assay by using L-Dopa as the substract. For melanin assay, I cultured the cells with my extracts, then extract the melanin from the cells and read the absorbance value using plate reader. I tried the melanin assay but the absorbance value that I get was very low...color produced very light. Still keep on trying......
Theoritically I know about the assays from reseach papers, but I don't know how to hands on carry out the experiments, I just keep on try an error...learn from mistakes. I am new to cell lysis and protein work...Research papers too brief, I need more detailed information for the assays...The more I read from journals, the more I am lost.