June 3, 015

Dear Murugadas

Personally I have never assayed these particular enzyme systems. But, you can use, as usually the case for other enzymes,  around 1 microgram per assay of 1 ml, if you work with (partially) purified samples. If you assay them in crude samples then around 10 micrograms could work. At the first time, it is required to assay their activity using different enzyme amounts in order to observe linear reaction progress (initial velocity) throughout a fixed reaction time.  The assay final volume depends on your luminometer plate (or cuvette) capacity, 0.2 to 0.5 ml is supposed to be sufficient. Note that initial velocity of enzyme depends on the assay conditions and the molecular activity of the assayed enzyme.

I hope this general comment is helpful

Thanks Prof Joseph,

I have not performed any enzyme assay earlier which made me to struck here, Since, it is luminometer plate i guess 0.2 ml will be sufficient. Yeah it is crude (whole animal) protein. I take 10 Microgram as per your suggestion.

Thanks again