ATP and Protein Quantification Protocol

ATP content was measured using the ATP bioluminescence assay kit CLS II (Roche, Heidelberg, Germany) as previously described (Shu et al., 2021 (link)). Protein content was measured using the Pierce BCA Assay kit. 20 μg of protein was run in duplicate in a white, flat-bottom 96-well microplate. Determination of free ATP was performed against an ATP standard curve. Luminescence was determined at 60 ms integration using a Synergy H1 Plate Reader (BioTek, Winooski, VT, United States).

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Shu D.Y., Frank S.I., Fitch T.C., Karg M.M., Butcher E.R., Nnuji-John E., Kim L.A, & Saint-Geniez M. (2022). Dimethyl Fumarate Blocks Tumor Necrosis Factor-Alpha-Driven Inflammation and Metabolic Rewiring in the Retinal Pigment Epithelium. Frontiers in Molecular Neuroscience, 15, 896786.

Publication 2022

CLS II Synergy H1 Plate Reader

Assay Bioluminescence assay Luminescence Protein

Corresponding Organization : Smith-Kettlewell Eye Research Institute

Other organizations : Cold Spring Harbor Laboratory

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

ATP content
Protein content

control variables

20 μg of protein was run in duplicate in a white, flat-bottom 96-well microplate
Luminescence was determined at 60 ms integration using a Synergy H1 Plate Reader (BioTek, Winooski, VT, United States)

controls

Determination of free ATP was performed against an ATP standard curve

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