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Radical reactions can occur either intermolecularly or intramolecularly. In an intermolecular radical reaction, a nucleophilic radical adds to an electrophilic alkene or vice versa. In such reactions, the radical and generally the alkene, which is also called the radical trap, are two different molecules. Additionally, for such intermolecular reactions to occur, the radical trap must be active, present in an excess concentration, and the radical starting material must have a weak...
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Updated: Dec 29, 2025

Rapid Scan Electron Paramagnetic Resonance Opens New Avenues for Imaging Physiologically Important Parameters In Vivo
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Intrinsic Buffer Hydroxyl Radical Dosimetry Using Tris(hydroxymethyl)aminomethane.

Addison E Roush1,2, Mohammad Riaz1, Sandeep K Misra1

  • 1Department of BioMolecular Sciences , University of Mississippi , Oxford , Mississippi 38677 , United States.

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|February 8, 2020
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Tris buffer acts as a novel hydroxyl radical dosimeter in Fast Photochemical Oxidation of Proteins (FPOP) experiments. This method allows for real-time monitoring and adjustments during protein footprinting analysis.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Chemical Biology

Background:

  • Fast Photochemical Oxidation of Proteins (FPOP) is a technique for protein surface footprinting using hydroxyl radicals.
  • Hydroxyl radical dosimeters are crucial for standardizing FPOP experiments due to variable radical production.
  • Previous FPOP studies in Tris buffer showed unexpected dosimetry behavior.

Purpose of the Study:

  • To investigate the behavior of Tris buffer under oxidative conditions in FPOP.
  • To determine if Tris can function as a hydroxyl radical dosimeter.
  • To explore the potential for real-time monitoring and adjustment in FPOP experiments.

Main Methods:

  • Utilizing laser flash photolysis of hydrogen peroxide to generate hydroxyl radicals.
  • Performing FPOP experiments with protein and adenine optical radical dosimetry in Tris buffer.
  • Detailed analysis of Tris buffer's oxidative behavior and signal response.

Main Results:

  • Tris buffer exhibits a novel gain-of-signal behavior when acting as an optical hydroxyl radical dosimeter.
  • This Tris-based dosimetry is suitable for inline, real-time monitoring.
  • The findings allow for dynamic adjustments to compensate for sample-specific quenching variations.

Conclusions:

  • Tris buffer can serve as an effective and novel hydroxyl radical dosimeter for FPOP.
  • The real-time monitoring capability of this Tris dosimeter enhances FPOP experimental reliability.
  • This discovery offers improved control and accuracy in protein footprinting studies.