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Mitotox complete oxphos activity assay panel

Manufactured by Abcam
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Sourced in United States

The MitoTox Complete OXPHOS Activity Assay Panel is a lab equipment product that measures the activity of the five complexes of the oxidative phosphorylation (OXPHOS) system in isolated mitochondria. It provides a comprehensive assessment of mitochondrial function.

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Fluostar optima 6, by BMG Labtech (1 mentions)

2 protocols using mitotox complete oxphos activity assay panel

1

Mitochondrial Complex Activity Assay

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The measurement of mitochondrial respiratory chain complex activity was performed by the MitoTox Complete OXPHOS Activity Assay Panel (# ab110419, Abcam) following the manufacturer's protocol 22 (link). In brief, each of the complexes was obtained from isolated mitochondria of the treated cells using highly specific monoclonal antibodies attached to 96-well microplates. Complex activity was determined by measuring the decrease in optical density per min as described in the manufacturer's protocol. Specified wavelengths (340 nm for complexes I and V, 550 nm for complexes III and IV, and 600 nm for complex II) were used to measure the absorbance.
Yi X., Guo W., Shi Q., Yang Y., Zhang W., Chen X., Kang P., Chen J., Cui T., Ma J., Wang H., Guo S., Chang Y., Liu L., Jian Z., Wang L., Xiao Q., Li S., Gao T, & Li C. (2019). SIRT3-Dependent Mitochondrial Dynamics Remodeling Contributes to Oxidative Stress-Induced Melanocyte Degeneration in Vitiligo. Theranostics, 9(6), 1614-1633.
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2

Ketamine's Impact on Mitochondrial Complexes

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The direct effect of ketamine on the electron transport chain complexes (I, II, IV and V) was measured using the MitoTox Complete OXPHOS Activity Assay Panel (Abcam, Cambridge, MA, USA) following the manufacturer’s protocol. Each of the complexes was obtained from isolated bovine heart mitochondria in their functionally active state using highly specific monoclonal antibodies attached to 96-well microplates. For each of the complexes treated with ketamine (31, 125, 500 μM), complex activity was determined by measuring the decrease in absorbance in milli-optical density per min at room temperature or 37°C, as described in the manufacturer’s protocol. Specified wavelengths (340 nm for complexes I and V, 600 nm for complex II, and 550 nm for complex IV) in kinetic mode (every min for 2 h) were used to measure the absorbance, using a FLUOstar OPTIMA-6 (BMG Labtech, Durham, NC, USA) microplate reader.
Ito H., Uchida T, & Makita K. (2015). Ketamine Causes Mitochondrial Dysfunction in Human Induced Pluripotent Stem Cell-Derived Neurons. PLoS ONE, 10(5), e0128445.
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