Citrate Synthase Activity Assay: A Colorimetric Assay to Measure Mitochondrial Citrate Synthase Activity in Drosophila Tissue Homogenate

Published: April 30, 2023

Abstract

Source: Wei, P., et al., A Colorimetric Assay of Citrate Synthase Activity in Drosophila Melanogaster. J. Vis. Exp. (2020)

This video describes a colorimetric assay to measure the mitochondrial citrate synthase activity assay present in the tissue homogenate of Drosophila.

Protocol

1. Colorimetric Citrate Synthase Activity Assay for D. melanogaster Collect ten adult flies for each sample. Collect at least triplicate samples for each genotype. Prepare 500 µL of ice-cold extraction buffer containing 20 mM HEPES (pH = 7.2), 1 mM EDTA, and 0.1% triton X-100 in a 1.5 mL test tube for each sample. Anesthetize adult flies with CO2 on an anesthesia pad and isolate the desired tissues. To isolate the adult fly thoraxes, for example, fix the fly t…

開示

The authors have nothing to disclose.

Materials

2-[4-(2-Hydroxyethyl)-1- piperazinyl]ethanesulfonic acid (HEPES)  Sigma-Aldrich  V900477
2-Amino-2-(hydroxymethyl)-1,3- propanediol (TRIZMA Base) Sigma-Aldrich V900483
Acetyl-CoA Sigma-Aldrich A2181
Dithio-bis-nitrobenzoic acid (DTNB) Sigma-Aldrich D8130
Ethylenediaminetetraacetic acid (EDTA) Sigma-Aldrich V900106
Oxaloacetate Sigma-Aldrich O4126
Pellet pestle Sangon F619072
Pellet pestle motor Tiangen OSE-Y10
Plate reader BioTek Eon
Protein BCA Assay kit Beyotime P0010S
Scissors WPI 14124
Triton X-100 Sangon A110694-0100

タグ

Play Video

記事を引用
Citrate Synthase Activity Assay: A Colorimetric Assay to Measure Mitochondrial Citrate Synthase Activity in Drosophila Tissue Homogenate. J. Vis. Exp. (Pending Publication), e21157, doi: (2023).

View Video