Overview

  • 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. 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

  • 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 1 x 15ml
    HRP Development Solution 1 x 12ml
  • Research areas
  • 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
    modifications
    The N-terminus is blocked.
  • Cellular localization
    Peroxisome.
  • Information by UniProt
  • Alternative names
    • CAT
    • CATA_HUMAN
    • Catalase
    • EC 1.11.1.6
    • MGC138422
    • MGC138424
    see all
  • Database links

Images

  • Example sample curves demonstrating the working range of the assay for a human and cultured cell line lysate (HepG2).
  • Example standard curve for the purified human catalase sample ab91026.
  • Example sample curves demonstrating the working range of the assay for a human and cultured cell line lysate (HepG2).
  • Example standard curve for the purified human catalase sample ab91026.
  • 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.

Protocols

References

This product has been referenced in:
  • Piao X  et al. 1-Deoxynojirimycin (DNJ) Ameliorates Indomethacin-Induced Gastric Ulcer in Mice by Affecting NF-kappaB Signaling Pathway. Front Pharmacol 9:372 (2018). Read more (PubMed: 29725297) »
  • Lespay-Rebolledo C  et al. Regionally Impaired Redox Homeostasis in the Brain of Rats Subjected to Global Perinatal Asphyxia: Sustained Effect up to 14 Postnatal Days. Neurotox Res 34:660-676 (2018). Read more (PubMed: 29959728) »
See all 4 Publications for this product

Customer reviews and Q&As

Answer

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