PMID: 2106829Mar 1, 1990Paper

A particle concentration fluorescence immunoassay for prostaglandin D synthase in the rat central nervous system

Archives of Biochemistry and Biophysics
F IslamO Hayaishi

Abstract

A solid phase, particle concentration fluorescence immunoassay (PCFIA) was developed for the measurement of prostaglandin (PG) D synthase in the 100,000g supernatant of various regions of the rat central nervous system. In this assay, the enzyme (in the range of 1-25 micrograms protein of brain supernatant or 1-100 ng of the purified enzyme) is attached to submicrometer carboxypolystyrene beads coated with polyclonal anti-rat brain PGD synthase IgG. The total particle-bound enzyme is assayed with fluorescein isothiocyanate (FITC)-conjugated monoclonal anti-PGD synthase IgG after incubation for 1 h. The optimum assay condition was obtained when carboxyl particles coated with ca. 500 micrograms/ml of polyclonal IgG at pH 5.0 and 5 micrograms/ml of FITC-IgG were used. No significant fluorescence was observed when FITC conjugates or carboxyl particles were prepared using IgG from nonimmunized rabbits. Heat treatment of the brain supernatant decreased the specific binding of the enzyme in parallel with the loss of enzyme activity, indicating that the denatured enzyme is not recognized by this assay method. The PGD synthase immunoreactivity was widely distributed in the brain regions and was highest in the paraflocculus. Although sli...Continue Reading

References

Dec 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·T ShimizuO Hayaishi
Feb 1, 1988·Archives of Biochemistry and Biophysics·M UjiharaO Hayaishi
Apr 3, 1986·Journal of Immunological Methods·J MayusM Nahm
Oct 15, 1987·The New England Journal of Medicine·J MiletichG Broze
Mar 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·R UenoO Hayaishi
Oct 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·R UenoO Hayaishi

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Citations

Aug 15, 1991·Archives of Biochemistry and Biophysics·F IslamO Hayaishi

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