Description
Key facts
Sample types:Cell culture extracts, Tissue Extracts,
Reacts with:Mouse, Rat, Human,
Assay type:Enzyme activity,
Range:1 - 1000 µg/mL,
Assay time:6h,
Assay Platform:Microplate reader
Product details:
Catalase Activity Assay Kit ab118184 is used to determine the relative specific activity (activity and quantity) of catalase in a sample. Capture antibodies specific for catalase are pre-coated in the microplate wells. The native enzyme is immunocaptured within the wells of the microplate; this removes all other enzymes.
The catalase assay buffer contains hydrogen peroxide which reacts with a substrate to make a luminescent product. Catalase functions rapidly to remove hydrogen peroxide from the solution and reduce the production of light. Therefore the light produced in each well is inversely proportional to the amount of catalase activity.
After catalase activity measurement the quantity of catalase is measured by adding an anti-catalase detector antibody to each well. After washing away unbound detector antibody, HRP-conjugated label specific for the detector antibody is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of catalase bound. The developing blue color is measured at 600 nm. Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.
Catalase assay protocol summary:
- add samples to wells
- incubate for 3 hrs at room temp
- wash with wash buffer
- add activity solution and hydrogen peroxide solution
- analyze with luminescence microplate reader for 30 min
- wash
- add detector antibody and incubate for 1 hr at room temp
- wash
- add HRP label and incubate for 1 hr at room temp
- wash
- add HRP development solution and analyze with microplate reader for 15 min in kinetic mode
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Catalase also known as CAT is an enzyme that catalyzes the decomposition of hydrogen peroxide into water and oxygen. This enzyme has a molecular weight of approximately 240 kDa and typically forms a tetramer. Catalase mainly resides in peroxisomes functioning as a peroxisome marker. It expresses abundantly in the liver kidneys and erythrocytes where it plays significant roles in cellular protection against oxidative damage. The presence of Catalase makes it an ideal candidate for use in peroxisome staining and peroxisome image analysis in research.
Biological function summary
Catalase contributes to antioxidant defense by breaking down hydrogen peroxide preventing cellular damage. In peroxisomes it works alongside other peroxisomal enzymes to maintain cell health and metabolic regulation. Catalase does not form a complex but interacts closely with other enzymes like superoxide dismutase which dismutates superoxide radicals into less harmful substances. Increased Catalase activity levels can be measured using Catalase activity kits and activity assays allowing us to learn about peroxisome function.
Pathways
Hydrogen peroxide removal by Catalase is vital in the reactive oxygen species (ROS) metabolic process and plays a part in the cellular response to oxidative stress. Catalase interacts with the glutathione peroxidase pathway safeguarding cells from oxidative stress-related damage. Superoxide dismutase works synergistically with Catalase transforming superoxide anions into hydrogen peroxide before its decomposition by Catalase. These activities highlight the essential role Catalase plays in protecting cells from oxidative stress damage.
Catalase relates to conditions like acatalasemia and diabetes. Acatalasemia a condition caused by a deficiency of Catalase increases the risk of developing diabetes and other oxidative stress-related diseases. Mutations in the CAT gene can lead to decreased Catalase activity contributing to the onset of these conditions. Additionally Catalase works with other proteins like glutathione peroxidase in mitigating the effects of oxidative stress with deficiencies potentially exacerbating complications in diabetes management.
