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Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells
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A fluorogenic dye activated by S-nitrosothiols.

Jia Pan1, Jonathan A Downing, Jeanne L McHale

  • 1Department of Chemistry, Washington State University, Pullman, WA, USA.

Molecular Biosystems
|August 12, 2009
PubMed
Summary
This summary is machine-generated.

A novel coumarin-phosphine dye activates its fluorescence when reacting with S-nitrosothiols. This discovery offers a new tool for detecting these important biological molecules.

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Area of Science:

  • Chemical Biology
  • Organic Chemistry
  • Fluorescent Probes

Background:

  • S-nitrosothiols (RSNOs) are crucial signaling molecules involved in various physiological and pathological processes.
  • Detecting and quantifying RSNOs in biological systems remains challenging due to their instability and low concentrations.

Purpose of the Study:

  • To develop a novel fluorescent probe for the sensitive detection of S-nitrosothiols.
  • To characterize the photophysical properties and reactivity of the new probe.

Main Methods:

  • Synthesis of a coumarin-phosphine dye.
  • Spectroscopic analysis (absorption, fluorescence) of the dye's response to S-nitrosothiols.
  • Evaluation of probe selectivity and sensitivity.

Main Results:

  • A coumarin-phosphine dye was successfully synthesized.
  • The dye exhibited quenched fluorescence that was activated upon reaction with S-nitrosothiols.
  • The probe demonstrated high sensitivity and selectivity for S-nitrosothiols over other reactive nitrogen species.

Conclusions:

  • The developed coumarin-phosphine dye serves as an effective "turn-on" fluorescent probe for S-nitrosothiols.
  • This probe offers a valuable tool for investigating the biological roles of S-nitrosothiols in real-time.