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Flow cytometric analysis of nitric oxide production in human neutrophils using dichlorofluorescein diacetate in the presence of a calmodulin inhibitor.

Publication ,  Journal Article
Rao, KM; Padmanabhan, J; Kilby, DL; Cohen, HJ; Currie, MS; Weinberg, JB
Published in: J Leukoc Biol
May 1992

Dichlorofluorescein (DCFH) oxidation assay measures hydrogen peroxide (H2O2), which is a derivative of superoxide anion. We found that a calmodulin antagonist, W-13, which is known to inhibit superoxide anion generation enhanced the capacity of human neutrophils to oxidize DCFH. To investigate this discrepancy we studied the role of nitric oxide (NO) in DCFH oxidation. Pure NO was capable of oxidizing DCFH, and the product formed had spectral properties identical to oxidized DCFH produced by H2O2. The arginine analog, NG-monomethyl-L-arginine (NMMA), which inhibits NO production, in combination with W-13 completely inhibited the stimulus-induced increase in DCFH oxidation. We conclude that the oxidation of DCFH in human neutrophils can occur by either H2O2 or NO.

Duke Scholars

Published In

J Leukoc Biol

DOI

ISSN

0741-5400

Publication Date

May 1992

Volume

51

Issue

5

Start / End Page

496 / 500

Location

England

Related Subject Headings

  • omega-N-Methylarginine
  • Sulfonamides
  • Solutions
  • Oxidation-Reduction
  • Nitric Oxide
  • Neutrophils
  • Immunology
  • Humans
  • Fluoresceins
  • Flow Cytometry
 

Citation

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Rao, K. M., Padmanabhan, J., Kilby, D. L., Cohen, H. J., Currie, M. S., & Weinberg, J. B. (1992). Flow cytometric analysis of nitric oxide production in human neutrophils using dichlorofluorescein diacetate in the presence of a calmodulin inhibitor. J Leukoc Biol, 51(5), 496–500. https://doi.org/10.1002/jlb.51.5.496
Rao, K. M., J. Padmanabhan, D. L. Kilby, H. J. Cohen, M. S. Currie, and J. B. Weinberg. “Flow cytometric analysis of nitric oxide production in human neutrophils using dichlorofluorescein diacetate in the presence of a calmodulin inhibitor.J Leukoc Biol 51, no. 5 (May 1992): 496–500. https://doi.org/10.1002/jlb.51.5.496.
Rao KM, Padmanabhan J, Kilby DL, Cohen HJ, Currie MS, Weinberg JB. Flow cytometric analysis of nitric oxide production in human neutrophils using dichlorofluorescein diacetate in the presence of a calmodulin inhibitor. J Leukoc Biol. 1992 May;51(5):496–500.
Rao, K. M., et al. “Flow cytometric analysis of nitric oxide production in human neutrophils using dichlorofluorescein diacetate in the presence of a calmodulin inhibitor.J Leukoc Biol, vol. 51, no. 5, May 1992, pp. 496–500. Pubmed, doi:10.1002/jlb.51.5.496.
Rao KM, Padmanabhan J, Kilby DL, Cohen HJ, Currie MS, Weinberg JB. Flow cytometric analysis of nitric oxide production in human neutrophils using dichlorofluorescein diacetate in the presence of a calmodulin inhibitor. J Leukoc Biol. 1992 May;51(5):496–500.

Published In

J Leukoc Biol

DOI

ISSN

0741-5400

Publication Date

May 1992

Volume

51

Issue

5

Start / End Page

496 / 500

Location

England

Related Subject Headings

  • omega-N-Methylarginine
  • Sulfonamides
  • Solutions
  • Oxidation-Reduction
  • Nitric Oxide
  • Neutrophils
  • Immunology
  • Humans
  • Fluoresceins
  • Flow Cytometry