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Catalase Activity Assay Kit ab118184 is used to determine the relative specific activity (activity and quantity) of catalase in a sample.

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Images

Functional Studies - Catalase Activity Assay Kit (Immunocapture) (AB118184), expandable thumbnail
  • Functional Studies - Catalase Activity Assay Kit (Immunocapture) (AB118184), expandable thumbnail
  • Functional Studies - Catalase Activity Assay Kit (Immunocapture) (AB118184), expandable thumbnail
  • Functional Studies - Catalase Activity Assay Kit (Immunocapture) (AB118184), expandable thumbnail
  • Functional Studies - Catalase Activity Assay Kit (Immunocapture) (AB118184), expandable thumbnail

Publications

Key facts

Sample types

Cell culture extracts, Tissue Extracts

Assay type

Enzyme activity

Reactive species

Mouse, Rat, Human

Range

1 - 1000 µg/mL

Assay time

6h

Associated Products

Select an associated product type

1 product for Alternative Product

Target data

Function

Catalyzes the degradation of hydrogen peroxide (H(2)O(2)) generated by peroxisomal oxidases to water and oxygen, thereby protecting cells from the toxic effects of hydrogen peroxide (PubMed:7882369). Promotes growth of cells including T-cells, B-cells, myeloid leukemia cells, melanoma cells, mastocytoma cells and normal and transformed fibroblast cells (PubMed:7882369).

Alternative names

What's included?

1 x 96 Tests
Components
10X Blocking Solution
1 x 10 mL
10X Detector Antibody
1 x 1.5 mL
10X HRP Label
1 x 1.5 mL
200X Luiminescent reagent
1 x 0.2 mL
20X Buffer
1 x 20 mL
55X Coupler
1 x 0.5 mL
60X Hydrogen Peroxide
1 x 0.5 mL
96-Well Microplate
1 x 1 Unit
Base Buffer
1 x 24 mL
Extraction Buffer (ab260490)
1 x 15 mL
HRP Development Solution
1 x 12 mL

Recommended products

Catalase Activity Assay Kit ab118184 is used to determine the relative specific activity (activity and quantity) of catalase in a sample.

Key facts

Sample types

Cell culture extracts, Tissue Extracts

Assay type

Enzyme activity

Reactive species

Mouse, Rat, Human

Range

1 - 1000 µg/mL

Assay time

6h

Assay Platform

Microplate reader

Precision

Intra assay

Sample

Sample 1

n

8

mean

-

SD

-

C.V.

7.1

Inter assay

Sample

Sample 1

n

4

mean

-

SD

-

C.V.

9.9

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

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

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Associated diseases and disorders

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.

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5 product images

  • Functional Studies - Catalase Activity Assay Kit (Immunocapture) (ab118184), expandable thumbnail

    Functional Studies - Catalase Activity Assay Kit (Immunocapture) (ab118184)

    Example relative specific activity – comparing the activity and quantity of purified erythrocyte catalase (Native human Catalase protein ab91026) and catalase from human hepg2 lysate. The HepG2 sample is determined to have a higher specific activity rate.

  • Functional Studies - Catalase Activity Assay Kit (Immunocapture) (ab118184), expandable thumbnail

    Functional Studies - Catalase Activity Assay Kit (Immunocapture) (ab118184)

    Example standard curve for the purified human catalase sample Native human Catalase protein ab91026.

  • Functional Studies - Catalase Activity Assay Kit (Immunocapture) (ab118184), expandable thumbnail

    Functional Studies - Catalase Activity Assay Kit (Immunocapture) (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 (Immunocapture) (ab118184), expandable thumbnail

    Functional Studies - Catalase Activity Assay Kit (Immunocapture) (ab118184)

    Example standard curve for the purified human catalase sample Native human Catalase protein ab91026.

  • Functional Studies - Catalase Activity Assay Kit (Immunocapture) (ab118184), expandable thumbnail

    Functional Studies - Catalase Activity Assay Kit (Immunocapture) (ab118184)

    Example sample curves demonstrating the working range of the assay for a human and cultured cell line lysate (HepG2).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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