Catalase enzyme assay protocol
Catalase assay flow chart
HPI activity was detected in all the isolates tested in the study. Introduction Catalase is a ubiquitous antioxidant enzyme that degrades hydrogen peroxide into water and oxygen 1. In addition, the developed method could measure the catalase activity in human cells, where the activity in LNCaP was higher than that in PC For the catalse assay, each test cuvette will need to be run one at a time, so do not prepare the next test cuvette until the run with the preceding cuvette is complete. Keep the samples on ice. Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue. Several studies have focused on these 2 catalases, as they play pivotal roles in protecting cells against the effects of oxidative stress 6.
Catalase activities of A laboratory strains of E. Figure 1: Image showing the height of foam developed in each test tube following the reaction of catalase with hydrogen peroxide, and the calibration curve plotted using the defined unit of catalase activity.
Furthermore, during quantitative analyses, although the reaction occurs spontaneously, the enzyme increases the reaction rate considerably. In conclusion, we have developed a simple and cost-effective assay and demonstrated its feasibility in determining the catalase status of bacterial and human cells.
All other chemicals were provided from standard commercial suppliers and were of analytical grade. Introduction Catalase is a ubiquitous antioxidant enzyme that degrades hydrogen peroxide into water and oxygen 1.
Simple spectrophotometric assay for measuring catalase activity in biological tissues
Nat Protoc 5 1 : The identified catalase activities instrumental measured activity were determined and plotted against the expected catalase activity at a series of dilutions of red blood cell homogenate Fig. Low concentrations of H2O2 are used to avoid bubbling. However, the activity of HPII varied among the isolates. Prepare fresh to each activity assay. Several studies have focused on these 2 catalases, as they play pivotal roles in protecting cells against the effects of oxidative stress 6. Dilute the lysate samples with PB. Following completion of the reaction, the height of O2-forming foam that remained constant for 15 min in the test tube was finally measured using a ruler. The authors will be requested to answer your questions at their earliest convenience. Catalase Activity Assay in Candida glabrata. A previous report has in fact demonstrated that a catalase-deficient mutant pathogen was more susceptible than its wild-type strain to the oxidative stress induced by hydrogen peroxide and immune cell attacks which involve hydrogen peroxide 2. Dilute the lysates or with sterile water.
Table 4 Analytical recovery of activity of catalase enzyme added to the reaction solution Full size table Conversion of hydrogen peroxide H2O2 to molecular oxygen and water H2O in a chemical reaction regulated by catalase was monitored by determining the absorbance of the carbonato-cobaltate III complex as a function of time as shown in Fig.
Methods Enzymol Finally, this assay comprises simple procedures, in that it involves only the mixing of the bacterial suspension, Triton X, and hydrogen peroxide solution, without any other procedures such as the preparation of bacterial extract.
Three surfactants produced similar results.
On the other hand, quantitative approaches focusing on careful measurements include colorimetric and spectrophotometric assays 910111213 The calibration curve plotted using the defined unit of catalase activity is shown in Fig.
based on 73 review