Dear Friends
Im searching a method for enzymatic assay of alkaline protease not using folin ciocalteu's phenol reagent. Could you give me a suggestion ?
You could use azocasein.
http://www.sigmaaldrich.com/catalog/product/sigma/a2765?lang=en®ion=US
I would like to recommend RP-HPLC-photometric method (please see file; Lysozyme by RP-HPLC), which is quantitative and reliable. Identifiction of products should be performed by using PDMD method, which is also quantitative and reliable (please see HepG2 fucoidan).
Direct photometric assay gives 4-fold lower values than HPLC-photometric method in the case of fluorescent-fucoidan determination (please see the Fig. 10 in the file SEC fucoidan determination). This is due to using photometer without purify, then curved calibration curve is obtained. Lambert-Beer's law is saying that light decreases according to atom or molecule in the light path; i.e., linear calibration line inserting to zero point can be obtainable if pure molecule is measured photometrically.
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