What does O Dianisidine do?
Product Description This product is a preweighed vial containing 50 mg of o-dianisidine dihydrochloride, which is a peroxidase substrate used in the quantitative determination of glucose in aqueous solutions such as serum, using a coupled enzyme system with glucose oxidase and peroxidase (PGO enzymes).
What is O Dianisidine dihydrochloride?
o-Dianisidine (dihydrochloride) is a colorimetric peroxidase substrate that has been used as a substrate to semi-quantitatively measure lactate, uric acid (Item No. 16219), and glucose. 1,2. It has also been used as a reagent for the detection of various metals, thiocyanates, and nitrites.
How is glucose determined using O Dianisidine?
A thin layer of GO is entrapped over an oxygen electrode via a semipermeable dialysis membrane. Glucose concentration is determined by measuring the amount of oxygen consumed by the enzyme reaction. A negative potential is applied to the platinum cathode for a reductive detection of the oxygen consumption [57,58].
Why did the reaction proceed slowly at 0 C?
Why did reaction proceed slowly at 0 deg. C? [Too cold and no energy] Enzymes cannot function at all because they are too far out of homeostasis (too cold). Not as much energy in the solution and energy is needed for reaction to occur.
What is peroxidase function?
Structure and Function Peroxidases are a group of enzymes that catalyze the oxidation of a substrate by hydrogen peroxide or an organic peroxide. Most peroxidases are able to oxidize a variety of substrates of widely different structures, varying from halide ions to hydroquinones to aromatic azo compounds.
How to measure o-dianisidine in a lab?
The oxidized O-dianisidine (yellow/orange colored compound) is measured at 430nm. Take 3.5mL phosphate buffer (pH 6.5) in a clean dry cuvette. Add 0.2mL enzymes extract and 0.1mL freshly prepared O-dianisidine solution. Bring the assay mixture to 28-30°C and then place the cuvette in the spectrophotometer set at 430nm.
How to do a peroxidase assay in a lab?
Bring the assay mixture to 28-30°C and then place the cuvette in the spectrophotometer set at 430nm. Then, add 0.2mL 0.2M H 2 O 2 and mix. Immediately start the stop watch. Read the initial absorbance and then at every 30sec intervals up to 3min. If the rate of increase is very high, repeat the assay with diluted extracts.
How is glucose oxidase and peroxidase prepared?
Method Reagent A mixed enzyme-oxygen acceptor reagent is prepared as follows : Glucose oxidase 125 mg., peroxidase 0-5 mg., and 05 ml, of 1 % o-dianisidine (in 95 % ethanol) are taken and made up to 100 ml. with 0.5 M NaH2PO4,2H2O (adjusted to pH 70 with NaOH) and filtered through paper.
Are there any non specific enzymes in peroxidase?
Peroxidase (POD) includes in its widest sense a group of specific enzymes such as NAD-Peroxidase, NADP-Peroxidase, fatty acid peroxidase etc. as well as a group of very non-specific enzymes from different sources which are simply known as POD (donor: H 2O 2-oxidoreductase 1.11.1.7).