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. 1985 Jul;78(3):614–618. doi: 10.1104/pp.78.3.614

Nitrate Reductase from Monoraphidium braunii

Immunocytochemical Localization and Immunological Characterization 1

Antonio Lopez-Ruiz 1,2,3, Jose Manuel Roldan 1,2,3, Jean Pierre Verbelen 1,2,3, Jesus Diez 1,2,3
PMCID: PMC1064785  PMID: 16664292

Abstract

Homogeneous nitrate reductase (EC 1.6.6.2) from Monoraphidium braunii was obtained by means of affinity chromatography in blue-Sepharose and gel filtration. After electrophoresis in polyacrylamide, gel slices containing pure nitrate reductase were disrupted and injected into previously unimmunized rabbits. The antiserum produced after several weeks was found to inhibit the different activities of nitrate reductase to a similar degree. Monospecificity of the antiserum was demonstrated by Ouchterlony double diffusion and crossed immunoelectrophoresis. The antibodies were purified by immunoabsorption to Sepharose-bound nitrate reductase.

The intracellular location of nitrate reductase in green algae was examined by applying an immunocytochemical method to M. braunii cells. Ultrathin frozen sections were first treated with immunopurified anti-nitrate reductase monospecific antibodies, followed by incubation with colloidal gold-labeled goat antirabbit immunoglobulin G as a marker. The enzyme was specifically located in the pyrenoid region of the chloroplast.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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