unknown enzyme concentration - (Oct/01/2012 )
Hi. Im an undergrad medical student currently doing Advance Medical Science. Im interested in determining Na/K-ATPase activity and Ca-ATPase activity in rat lens.
Regarding
The question is, how to convert amount of the 2 enzymes (pyruvate kinase and lactate dehydrogenase) from microgram into concentration or activity unit?
" Aliquots of homogenate were assayed enzymatically for total ATPase activity in 1 ml reaction mixture containing 100 mmol/l NaCl,
10 mmol/l KCl, 2.5 mmol/l MgCl 2 , 1 mmol/l Tris-ATP, 1 mmol/l phosphoenolpyruvate,30 mmol/l imidazole-HCl buffer, pH 7.3, 0.15 mmol/l NADH,
here is an attachment of my flowchart regarding this assay.
your enzyme concentration, in the assay, would be 50ug/ml ldh and 30ug/ml pk. you can get the units from the spec sheet that comes with the enzyme (or you can go to the supplier's website and look up the specs for the lot#).
but, i'm not sure that that is what you meant to ask, is it?