What is the extinction coefficient for NADH?
= 6220 M-1cm-1
Use extinction coefficient for NADH (ε340 = 6220 M-1cm-1) to determine rate of ADP production (NADH loss). Remember that the path length of the stopped-flow cuvette is 0.2 cm.
What is the extinction coefficient for NADPH?
6.22 mM-1 cm-1
The extinction coefficient of NADPH is 6.22 mM-1 cm-1, which can be used to directly quantify the formation and at the end the total product formed during the reaction, assuming the concentration of substrate S is limited in solution and NADP+ is available in excess.
What is the extinction coefficient ε of NADH at the wavelength we monitor the absorbance at?
6.22 x 103 cm-1M-1
NADH has an absorption band centered at 340 nm with an extinction coefficient of 6.22 x 103 cm-1M-1, while NAD+ has no absorbance at this wavelength.
What is the molar extinction coefficient of NADH at 340 nm?
NADH has an extinction coefficient of 6220 M-1 cm-1 at 340nm. The extinction coefficient for NADH is 6220 M −1 − 1 cm −1 − 1 at 340 nm.
What is the extinction coefficient of NADH at 340 nm?
6.22 l/mmol/cm
The molar extinction coefficient for NADH or NADPH at 340 nm is 6.22 l/mmol/cm, which, according to the Beer–Lambert law, means that a solution of 0.1 mM would have an optical density equal to 0.622 through a 1-cm light path.
How is Nadph production measured?
The amount of NADP+ produced by the reaction is quantified by comparison to known concentrations of NADP+ standards. The NAD+ kinase activity is calculated by subtracting the amount of NADP+ produced by the reaction from the amount of NADP+ produced without the addition of the tissue lysate.
What is milli absorbance unit?
Absorbance (Abs) in chemistry is a measure of the attenuation of the transmitted radiant power in a material. It is a numerical value that shows the capacity of a substance to absorb light of a specified wavelength. Milli Absorbance (mAbs) is one-thousandth of an absorbance unit.
What is the molar absorptivity of NADH at 340 nm?
The molar absorptivity of NADH at 340 nm has been determined by an indirect procedure in which high-purity glucose is phosphorylated by ATP in the presence of hexokinase, coupled to oxidation of the glucose-6-phosphate by NAD+ in the presence of glucose-6-phosphate dehydrogenase.
What is the best way to measure the concentration of NADH?
NADH can be readily measured in a spectrophotometer at 340nm. This protocol details the use of the Glucose (HK) Assay Kit from Sigma (product code GAHK-20).
How much is 6317 mol-1 cm-1 at 25 degrees?
The average value from 85 independent determinations is 6317 liter mol-1 cm-1 at 25 degrees C and pH 7.8.