Quantitative Glutathione Measurement in DRG Neurons

Assessment of reduced intracellular glutathione (GSH) was with Thioltracker Violet (Life Technologies) cell-permeant thiol-reactive fluorescent probe as described [46 (link),47 (link)]. DRG neurons were seeded in black wall 96-well plates (10⁴/well). After treatments, cultures were washed with PBS. Freshly prepared 10 μM Thioltracker Violet in DPBS was added for 30 min/37°C incubation in the dark. Fluorescence was measured using the FL200 Tecan (excitation at 415 nm/emission 530 nm). Four sets of independent experiments were carried out and mean±SEM values were calculated.

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Gorgun M.F., Zhuo M, & Englander E.W. (2016). Cisplatin Toxicity in Dorsal Root Ganglion Neurons is Relieved by Meclizine via Diminution of Mitochondrial Compromise and Improved Clearance of DNA Damage. Molecular neurobiology, 54(10), 7883-7895.

Publication 2016

Thioltracker Violet

After treatments Cell Fluorescence Fluorescent probe Intracellular Neurons Reduced glutathione Thiol Violet

Corresponding Organization :

Other organizations : The University of Texas Medical Branch at Galveston

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Variable analysis

independent variables

Treatments

dependent variables

Reduced intracellular glutathione (GSH) levels

control variables

DRG neurons seeded in black wall 96-well plates (10^4/well)
Cultures were washed with PBS
Freshly prepared 10 μM Thioltracker Violet in DPBS was added for 30 min/37°C incubation in the dark
Fluorescence was measured using the FL200 Tecan (excitation at 415 nm/emission 530 nm)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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