controls for enzyme assay - (Aug/23/2006 )
I'm working on a project investigating whether a complement component can regulate an enzyme; in one experiment i want to overexpress complement, then assay the enzyme levels. i'm not sure what controls to use for this, in order to demonstrate that the effect is specific for this enzyme, and not a global effect. are there any sort of "standard enzymes" that can be used as controls, ie complement expression should have no effect on them? i haven't found any good examples in the literature yet, and the only things i've thought of are glycolitic enzymes, or maybe ATPase. Does anyone have any suggestions?
ps - i'll be using hek293T cells
Since complement is involved with immune responses try enzymes involved with metabolism or even structural ones: Alkaline phosphatase, lipase, dehydrogenases in cortisol metabolism. Maybe even alanine transferase or aspartate transferase. Mind you, I always feel it's so complicated looking at immunology what with cytokines etc. You just don't know what is being invoked. Check the acute phase response with whatever ez you go for. I've got a notion that glycolysis is influenced by the immune response but cant remember the details.