Quantifying Mitochondrial Aconitase Activity

Relative mitochondrial aconitase activity was quantified using the colorimetric Aconitase Activity Assay Kit from Sigma (#MAK051), following the manufacturer’s instructions as previously described (Martelli et al., 2020 (link)). A total of six biological replicates (25 whole larvae per replicate) were exposed to 2.5 ppm spinosad for 2 hr, whilst six control replicates (25 whole larvae per replicate) were exposed to DMSO for 2 hr. Absorbance was measured at 450 nm in a FLUOstar OPTIMA (BMG LABTECH) microplate reader using the software OPTIMA and normalized to sample weight. The data were analyzed using one-way ANOVA followed by Tukey’s honestly significant difference test.

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Martelli F., Hernandes N.H., Zuo Z., Wang J., Wong C.O., Karagas N.E., Roessner U., Rupasinghe T., Robin C., Venkatachalam K., Perry T., Batterham P, & Bellen H.J. (2022). Low doses of the organic insecticide spinosad trigger lysosomal defects, elevated ROS, lipid dysregulation, and neurodegeneration in flies. eLife, 11, e73812.

Publication 2022

Aconitase Activity Assay Kit FLUOstar OPTIMA

Aconitase Anova Assay Biological Colorimetric Dmso Larvae Mitochondrial Replicate Spinosad

Corresponding Organization :

Other organizations : University of Melbourne, Baylor College of Medicine, The University of Texas Health Science Center at Houston, Neurological Research Institute, Howard Hughes Medical Institute

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Variable analysis

independent variables

Exposure to 2.5 ppm spinosad for 2 hr

dependent variables

Relative mitochondrial aconitase activity

control variables

Exposure to DMSO for 2 hr

controls

Negative control: Exposure to DMSO for 2 hr

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