BIOC 2580 Lecture 14: Day 14: Enzyme assays [Feb 08]
10 Feb 2017
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Measure rates: rate of disappearance of reactant or appearance of product specific enzyme substrate(s) product(s) catalase. 200-400 nm larger chromophores absorb at longer wavelengths. Most spectrophotometers read directly in absorbance units the absorbance of a sample is a measure of its concentration absorbance is proportional to sample concentration (beer"s law) and sample thickness (lambert"s law) absorbance a = c l. Beer-lambert law c = sample concentration in mol/l. = sample thickness in cm, usually 1. 00 cm. = the extinction coefficient, a constant for any given substance for nadh, = 6200 l/mol cm at 340 nm sample reaction using absorbance: lactate dehydrogenase reaction showed decrease in absorbance of. 0. 357 at 340 nm due to conversion of nadh and nad + sample is 1. 00 cm thick for nadh, = 6200 l/mol cm at 340 nm decrease in [nadh] = a / ( x l. = 5. 76x10 -5 mol/l quantitative description of enzyme catalysis.
