Catalase Activity Assay Kit (Immunocapture) (ab118184)
Key features and details
- Assay type: Enzyme activity
- Platform: Microplate reader
- Assay time: 6 hr
- Sample type: Cell culture extracts, Tissue Extracts
Overview
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Product name
Catalase Activity Assay Kit (Immunocapture)
See all Catalase kits -
Precision
Intra-assay Sample n Mean SD CV% 1 8 7.1% Inter-assay Sample n Mean SD CV% 1 4 9.9% -
Sample type
Cell culture extracts, Tissue Extracts -
Assay type
Enzyme activity -
Range
1 µg/ml - 1000 µg/ml -
Assay time
6h 00m -
Species reactivity
Reacts with: Mouse, Rat, Human -
Product overview
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 -
Platform
Microplate reader
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Blocking Solution 1 x 10ml 10X Detector Antibody 1 x 1.5ml 10X HRP Label 1 x 1.5ml 200X Luiminescent reagent 1 x 0.2ml 20X Buffer 1 x 20ml 55X Coupler 1 x 0.5ml 60X Hydrogen Peroxide 1 x 0.5ml 96-Well Microplate 1 unit Base Buffer 1 x 24ml Extraction Buffer (ab260490) 1 x 15ml HRP Development Solution 1 x 12ml -
Research areas
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Function
Occurs in almost all aerobically respiring organisms and serves to protect cells from the toxic effects of hydrogen peroxide. Promotes growth of cells including T-cells, B-cells, myeloid leukemia cells, melanoma cells, mastocytoma cells and normal and transformed fibroblast cells. -
Involvement in disease
Defects in CAT are the cause of acatalasia (ACATLAS) [MIM:115500]; also known as acatalasemia. This disease is characterized by absence of catalase activity in red cells and is often associated with ulcerating oral lesions. -
Sequence similarities
Belongs to the catalase family. -
Post-translational
modificationsThe N-terminus is blocked. -
Cellular localization
Peroxisome. - Information by UniProt
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Alternative names
- Cas1
- CAT
- CATA_HUMAN
see all -
Database links
- Entrez Gene: 847 Human
- Entrez Gene: 12359 Mouse
- Entrez Gene: 24248 Rat
- Omim: 115500 Human
- SwissProt: P04040 Human
- SwissProt: P24270 Mouse
- SwissProt: P04762 Rat
- Unigene: 502302 Human
see all
Images
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Functional Studies - Catalase Activity Assay Kit (ab118184)Example relative specific activity – comparing the activity and quantity of purified erythrocyte catalase (ab91026) and catalase from human hepg2 lysate. The HepG2 sample is determined to have a higher specific activity rate.
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Functional Studies - Catalase Activity Assay Kit (ab118184)Example standard curve for the purified human catalase sample ab91026. -
Functional Studies - Catalase Activity Assay Kit (ab118184)Example sample curves demonstrating the working range of the assay for a human and cultured cell line lysate (HepG2). -
Functional Studies - Catalase Activity Assay Kit (ab118184)Example standard curve for the purified human catalase sample ab91026. -
Functional Studies - Catalase Activity Assay Kit (ab118184)Example sample curves demonstrating the working range of the assay for a human and cultured cell line lysate (HepG2).
