Oxidative Stress and DNA Damage Assessment

Thiobarbituric acid reactive substances (TBAR) level was examined to reflect the production of toxic aldehyde resulting from oxidative fatty acyl degradation, the malondialdehyde (MDA). The detailed protocol was described previously [45 (link), 46 (link)].
γ-H2AX assay of DNA damages. ARPE-19 cells were fixed in ice-cold ethanol, which were then incubated with a mouse monoclonal anti-γ-H2AX antibody (Santa Cruz Biotech). Afterwards, the FITC-conjugated anti-mouse secondary antibody (Santa Cruz) was then added. Cells were then subjected to FACS assay to determine the γ-H2AX percentage, reflecting DNA damage intensity [47 (link)].

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Li C., Yan K., Wang W., Bai Q., Dai C., Li X, & Huang D. (2017). MIND4-17 protects retinal pigment epithelium cells and retinal ganglion cells from UV. Oncotarget, 8(52), 89793-89801.

Publication 2017

FITC-conjugated anti-mouse secondary antibody

Aldehyde Anti antibody Assay Cells Cold Dna damage Ethanol Fitc Malondialdehyde Monoclonal antibody Mouse Thiobarbituric acid reactive substances

Corresponding Organization :

Other organizations : Huaian First People’s Hospital, Nanjing Medical University

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Protocol cited in 9 other protocols

Variable analysis

independent variables

Oxidative fatty acyl degradation

dependent variables

Thiobarbituric acid reactive substances (TBAR) level
Malondialdehyde (MDA) level
γ-H2AX percentage (reflecting DNA damage intensity)

control variables

Not explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

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