An advanced oxidation protein products (AOPP) assay, which uses Chloramine-T as a marker of oxidative protein damage, was conducted on the plasma. The AOPP assay kit (ab242295; Abcam), was used in accordance with manufacturer's instructions. In summary, treated and non-treated plasma samples were diluted in PBS (1:10). 200 µL diluted plasma (n = 3), and standards of Chloramine-T (0-100-µM), were transferred to a 96-well plate and 10 µL of Chloramine Reaction Initiator was added to all wells. The plate was shaken for 5 min (120 rpm, room temperature) and 20 µL Stop Solution was added. Absorbance at 340 nm was measured, and AOPP levels expressed as µM of Chloramine-T equivalents, with values corrected for dilution factor.
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Quantifying Oxidative Protein Damage
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Corresponding Organization :
Other organizations : The Robertson Trust, University of Strathclyde, Center for Biologics Evaluation and Research
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Variable analysis
independent variables
- Plasma treatment (treated vs. non-treated)
- AOPP levels (expressed as μM of Chloramine-T equivalents)
- Dilution of plasma samples (1:10 in PBS)
- Chloramine-T standards (0-100 μM)
- Shaking time (5 min, 120 rpm, room temperature)
- Measurement of absorbance at 340 nm
- Positive control: Chloramine-T standards (0-100 μM)
- Negative control: Not explicitly mentioned
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