What is peroxidase assay?
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).
How do you calculate peroxidase in plants?
The enzyme activity can be calculated according to the following equation: Enzyme activity (Units/L) = (ΔAbs × Total assay volume) / ( Δt x ε x l x Enzyme sample volume).
Which substrate is used for assay of peroxidase?
This procedure is for the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate.
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.
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 is the purpose 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 the function of peroxidase?
Which is used as a substrate for the assay of peroxidase?
Guaiacol is used as substrate for the assay of peroxidase. The resulting oxidized (dehydrogenated) guaiacol is probably more than one compound and depends on the reaction conditions. The rate of formation of guaiacol dehydrogenation product is a measure of the POD activity and can be assayed spectrophotometrically at 436nm.
How can I calculate ascorbate peroxidase activity?
Ascorbate peroxidase was assayed by recording the decrease in optical density due to ascorbic acid at 290 nm . The 3 ml reaction mixture contained 50 mM potassium phosphate buffer (pH 7.0), 0.5 mM ascorbic acid, 0.1 mM EDTA, 0.1 mM H2O2, enzyme and water. The reaction was run for 5 min at 25 °C.
How is the rate of formation of guaiacol measured?
The rate of formation of guaiacol dehydrogenation product is a measure of the POD activity and can be assayed spectrophotometrically at 436nm. Dissolve 240mg guaiacol in water and make up to 100mL. It can be stored frozen for many months. » Hydrogen peroxide solution (0.042% = 12.3mM). Dilute 0.14mL of 30% H 2 O 2 to 100 mL with water.
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).