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Mitochondrial Stress Test Complete Assay Kit (ab232857)

  • Datasheet
  • SDS
  • Protocol Booklet
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Recommended plate map.
  • Signal Profiles of HepG2 oxygen consumption obtained using the Mitochondrial Stress Test Complete Assay.
  • Full characterization of mitochondrial function using the Mitochondrial Stress Test Complete Assay.
  • Basal Respiration and Maximal Respiration in HepG2 cells measured in different nutrient conditions.

Key features and details

  • Assay type: Cell-based
  • Detection method: Fluorescent
  • Platform: Microplate reader
  • Sample type: Adherent cells, Purified mitochondria, Suspension cells, Tissue

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Overview

  • Product name

    Mitochondrial Stress Test Complete Assay Kit
    See all Extracellular Oxygen Consumption kits
  • Detection method

    Fluorescent
  • Sample type

    Tissue, Adherent cells, Suspension cells, Purified mitochondria
  • Assay type

    Cell-based
  • Assay duration

    Multiple steps standard assay
  • Product overview

    Mitochondrial Stress Test Complete Assay Kit (ab232857) (HS Method) is a highly flexible 96- or 384-well fluorescence plate reader-based approach, for the direct, real-time analysis of cellular respiration and mitochondrial function.


    The easy-to-use assay allows measurement of extracellular oxygen consumption rates (OCR) with whole cell populations (both adherent and suspension cells), isolated mitochondria, permeabilized cells and a wide range of 3D cultures including: tissues, small organisms, spheroids, scaffolds and matrixes. The assay is also suitable for measurement of isolated enzymes, bacteria, yeasts and molds.


    The Extracellular O2 probe is chemically stable and inert, water-soluble and cell impermeable, making it the ideal and scalable mix-and-measure reagent for use in a wide range of cell culture conditions - all measured using a fluorescence plate-reader. In this assay, Extracellular O2 probe is quenched by O2, through molecular collision, and thus the amount of fluorescence signal is inversely proportional to the amount of extracellular O2 in the sample. Rates of oxygen consumption are calculated from the changes in fluorescence signal over time.


    The reaction is non-destructive and fully reversible (neither Extracellular O2 probe nor O2 are consumed), facilitating measurement of time courses and drug treatments.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 x 96 tests
    Antimycin A 1 x 3µg
    Extracellular O2 probe 1 vial
    FCCP 1 x 4µg
    Glucose Oxidase 1 x 112.5µg
    HS Mineral Oil 1 x 15ml
    Oligomycin 1 x 10µg

Associated products

  • Related Products

    • Extracellular Oxygen Consumption Reagent (ab197242)
    • Extracellular Oxygen Consumption Assay (ab197243)
    • Mitochondrial Stress Test Companion Assay (ab243390)
    • Mineral Oil High sensitivity (ab243855)

Images

  • Recommended plate map.
    Recommended plate map.

    Using this plate map, up to 7 different conditions can be tested simultaneously in triplicate across all treatment conditions.

  • Signal Profiles of HepG2 oxygen consumption obtained using the Mitochondrial Stress Test Complete Assay.
    Signal Profiles of HepG2 oxygen consumption obtained using the Mitochondrial Stress Test Complete Assay.

    Slopes (m) calculated from the linear portion of these signal profiles are reflective of the OCR under the conditions imposed by the kit components and are used to determine the key characteristics of aerobic respiration.

  • Full characterization of mitochondrial function using the Mitochondrial Stress Test Complete Assay.
    Full characterization of mitochondrial function using the Mitochondrial Stress Test Complete Assay.

    a) The OCR (m) from the linear portion of the signal profile from each Stress Test condition reflect Basal Respiration, Non ATP-Coupled Oxygen Consumption, Maximal Respiration and Non Respiratory Oxygen Consumption. ATP-Coupled Oxygen Consumption and Spare Respiratory Capacity are calculated from these values.
    b) The contribution of each of these discrete metabolic processes to the Maximal Respiration can be conveniently visualized producing a detailed picture of aerobic metabolism.

  • Basal Respiration and Maximal Respiration in HepG2 cells measured in different nutrient conditions.
    Basal Respiration and Maximal Respiration in HepG2 cells measured in different nutrient conditions.

    In low glucose medium, Spare Respiratory Capacity is low, but significantly increases in the presence of high glucose concentrations or 150 µM Oleate, due to the provision of reducing equivalents to fuel ETC activity.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (0)

    Publishing research using ab232857? Please let us know so that we can cite the reference in this datasheet.

    ab232857 has not yet been referenced specifically in any publications.

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