I am focusing on PPAR-Alpha expression by estimating various enzyme activity (CPT, FS, FAS, ME, G6PD, ACAT etc.) from Rat Liver homogenates..
I don't have the exact procedure for estimation of these enzymes..
Hi Bhavesh, I am not sure whether I understood your question correctly: PPARa is in first line transcription factor, thus its activity should be reflected in the measurments of the levels of ist downstream target gene expression, i.e. ACOX, CYP4A11, CPT, PDK4 and many others by using usual quantitative RT-PCR. Assessement of the activity of the enzymes you named will not nessesarily be the indicators of PPARa activity since enzymatic activity of CPT, FS, FAS, etc can be additionaly regulated via plenty of other factors. Correct me if I misunderstood your question and Good Luck!
Thank you Madam, for your kind reply.
You are correct, we are planning to check few downstream targets of PPARa but I am interested to various hepatic enzyme activitie like CPT, Hepatic beta oxidation, G6PD,ME, FAS, HmG-CoA reductase and LPL etc. by spectrophotometric method.
Can you suggest me such conventional methods for estimation of these enzyme??
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