<原著>In situ hybridizationを用いたβ-adrenergic receptorの分子レベルでの検討

概要

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A new approach is presented to reveal the location of β-adrenergic receptor in microscopic preparation. The method is based on the application of the probes, synthesized β-adrenergic receptor oligonucleotide sequence. After "in situ hybridization", the DNP labelled probes were recognized by antibodies and visualized immunocytochemically. This procedure was optimized on the frozen sections of fresh guinea pig lungs. Staining was indicated in β-adrenergic receptor DNA on the lung tissue. As the negative control PBR 322 DNA was used. The positive control was hybrids between promyelocytic leukemia cells (HL-60 cells) and c-myc DNA. Further, for c-DNA encoding β-adrenergic receptor, the β-adrenergic receptor protein from human placenta was purified by sequential Sephadex-alprenolol affinity chromatography. Antibodies against the receptor protein were produced by in vitro immunization on BALB/C mouse spleen cells. The m-RNA from human placenta was prepared by oligo (dT) cellulose column chromatography.
近畿大学の論文
1988-03-25