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Related Concept Videos

Nitric Oxide Signaling Pathway01:28

Nitric Oxide Signaling Pathway

Nitric oxide (NO), an inorganic gas, acts as a potent second messenger in most animal and plant tissues. NO diffuses out of the cells that produce it and enters the neighboring cells to generate a downstream response. NO synthase (NOS) catalyzes NO production by the deamination of the amino acid arginine. There are three isoforms of NOS. Endothelial cells have endothelial NOS (eNOS), nerve and muscle cells have neuronal NOS (nNOS), and macrophages produce inducible NOS (iNOS) upon exposure to...

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Analytical Techniques for Assaying Nitric Oxide Bioactivity
11:28

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Published on: June 18, 2012

An improved histochemical method for measuring nitric oxide synthase activity.

J Alaghband-Zadeh1, S Mehdizadeh, A O'Farrell

  • 1Endocrine Laboratory, Department of Clinical Chemistry, Imperial College of Science, Technology & Medicine (Charing Cross Campus), London, UK.

Cell Biochemistry and Function
|August 19, 1999
PubMed
Summary

A new method improves nitric oxide synthase (NOS) measurement in tissue sections. This technique enhances reagent preparation and activity measurement, ensuring all NOS activity is retained within the section for accurate results.

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Area of Science:

  • Biochemistry
  • Histochemistry
  • Enzyme Assays

Background:

  • Quantitative histochemical methods for measuring nitric oxide synthase (NOS) activity previously resulted in significant loss of enzyme activity.
  • Approximately 15% of NOS was lost from tissue sections into the reaction medium during prior measurement procedures.

Purpose of the Study:

  • To describe an improved quantitative histochemical method for measuring NOS activity in tissue sections.
  • To address the issue of enzyme loss during measurement by modifying reagent preparation and the assay procedure.

Main Methods:

  • The study details modifications to the preparation of the active reagent, lead ammonium citrate/acetate (LACA).
  • An improved procedure for measuring NOS activity within tissue sections was developed and implemented.

Main Results:

  • The revised method involves optimized LACA reagent preparation.
  • The new NOS activity measurement procedure effectively retains all measurable NOS activity within the tissue section.
  • This contrasts with previous methods where significant enzyme loss occurred.

Conclusions:

  • The developed method offers a more accurate and reliable way to quantify NOS activity in tissue sections.
  • By minimizing enzyme loss, the new technique provides a more complete assessment of NOS activity.
  • This advancement is crucial for histochemical studies involving nitric oxide synthase.