What does O Dianisidine enzyme reagent test for?
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 the function of peroxidase?
Factors Effecting the Rate of Peroxidase Reactions Peroxidase is an enzyme found in a wide variety of organisms, from plants to humans to bacteria. Its function is to break down hydrogen peroxide (H2O2), which is one of the toxins produced as a byproduct of using oxygen for respiration.
What is peroxidase assay?
Abcam’s Peroxidase Assay Kit (ab155895) provides a convenient colorimetric and fluorometric means to measure the peroxidase activity in biological samples. The resorufin is quantified by colorimetric (λmax = 570nm) or fluorometric methods (Ex/Em = 535/587 nm).
Is Dianisidine dihydrochloride a dye?
Benzidine derivatives, such as 3,30-dimethylbenzidine and 3,30-dimethoxybenzidine (o-dianisidine) (Figure 25), are used as dyes or intermediates for dyestuffs or pigments (e.g., Trypan Blue, Acid Red 14, Direct Blue 1, 8, 15, 76, 98, 218, and Pigment Orange 16), coatings, plastics, or in chemical studies.
What is the importance of peroxidase in blood?
Peroxidases play an important role in innate immunity and in a number of physiologically important processes like apoptosis and cell signaling. Unfavorable excessive peroxidase activity is implicated in oxidative damage of cells and tissues, thereby initiating the variety of human diseases.
Does peroxidase produce oxygen?
The rate of oxygen production by peroxidases was greatly enhanced by the addition of ABTS [2,2′-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid)] or chlorpromazine, both of which are oxidized to cation radicals by peroxidases.
Why is the activity of peroxidase and catalase in plant materials of so much concern?
Higher peroxidase and catalase activities indicate more intensive production of toxic hydrogen peroxide evolved by reactions of different type in green plants in comparison with etiolated ones. Significantly lower activity of peroxidases in etiolated plants could be related to a lower degree of lignification.
What is peroxidase activity of hemoglobin?
As a hemoprotein, hemoglobin (Hb) can, in the presence of H2O2, act as a peroxidase. For stroma-free Hb this regulation is lost, and the potential for Hb to become a peroxidase is high and further increased by inflammatory cells generating superoxide.
What factors affect peroxidase activity?
The results of this experiment demonstrated that the performance of hydrogen peroxidase was greatly affected by all three factors, temperature, concentration level, and pH. Of the conditions we tested, this enzyme was most active at a temperature of 38°C, a pH of 7, and at the highest concentration tested.
How is laccase activity calculated?
For determination of laccase activities you mix your enzyme solution with an ABTS stock solution at a defined pH (use a buffer solution like e.g. McIlvaine phosphate citrate buffer). ABTS concentration of the mixture should be 0.2 mM. Then you measure absorbance over time at 420 nm.
What is O Dianisidine dihydrochloride?
Product Description 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 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 make a peroxidase enzyme assay?
Vortex briefly and let settle before use. Store in amber bottle or wrapped in foil. Substrate: Prepare 0.3% H 2 0 2 solution in deionized or distilled water. Prior to use dilute with Buffer to a final concentration of 0.003%. Add 0.05 mls of Dye to 6.0 ml of substrate and vortex.
How to calculate enzyme activity of horseradish peroxidase?
One unit of activity is the amount of enzyme to decompose 1 µmole of peroxide/minute at 25 degrees Celsius. 11.3 x 10 3 cm -1 is the molar absorbance of H 2 O 2. Questions? Lost your password? If your country not listed above please click here.
How does the staining of dianisidine take place?
The gels or cross sections are first incubated in a buffered solution containing 100 m M KBr or 100 m M KI, and 10 m M o -dianisidine, and the staining is started by addition of 2 m M H 2 O 2. The formed HOBr or HOI will oxidize the organic dye resulting in polymerization and formation of a brown precipitate.