Determination of Brain Enolase Isozymes with an Enzyme Immunoassay at the Level of Single Neurons
抄録
<jats:p><jats:bold>Abstract</jats:bold> Ultrasensitive enzyme immunoassay systems for the assay of rat brain enolase isozymes (<jats:italic>αα</jats:italic>, <jats:italic>αγ</jats:italic>, and <jats:italic>γγ</jats:italic> forms) were prepared by use of β‐<jats:sc>d</jats:sc>‐galactosidase from <jats:italic>Escherichia coli</jats:italic> as label and the purified rabbit antibodies to <jats:italic>αα</jats:italic> and <jats:italic>γγ</jats:italic> enolases. The antibodies were purified from the immunoglobulin G (IgG) fractions of antisera by immunoaffinity chromatography with a column of the corresponding antigen‐coupled Sepharose. Sandwich‐type immunoassay systems with the galactosidase‐labeled antibody Fab’fragments and the antibody F(abapos;)<jats:sub>2</jats:sub>‐immobilized polystyrene beads could determine amounts as small as 1 amol (10<jats:sup>−18</jats:sup> mol) of each isozyme. Purkinje cell bodies picked up from the bulk‐separated fraction by means of a nylon loop were subjected to the assay at the level of single cells. In contrast to previous report, this neuron contained not only the <jats:italic>γγ</jats:italic> but also the <jats:italic>αγ</jats:italic> and <jats:italic>αα</jats:italic> enolases at a level of amol per cell body, although the concentration of <jats:italic>γγ</jats:italic> was the highest. Immunohistochemical experiments on the cerebellum with the peroxidase‐labeled antirabbit IgG antibody and the unlabeled antibody method confirmed the above results, and indicated that both <jats:italic>α</jats:italic> and γ subunits of the enolase were stained intensely in axons.</jats:p>
収録刊行物
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- Journal of Neurochemistry
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Journal of Neurochemistry 37 (4), 998-1005, 1981-10
Wiley
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詳細情報 詳細情報について
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- CRID
- 1362544421239095680
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- NII論文ID
- 30014579023
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- ISSN
- 14714159
- 00223042
- http://id.crossref.org/issn/00223042
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