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A Novel Amplification Method of Nonradioactive In Situ Hybridization Signal for Specific RNA with Biotinylated Tyramine
http://hdl.handle.net/10069/23455
http://hdl.handle.net/10069/23455798e54b1-f0c7-42a9-b27a-51e9be2a4f2e
名前 / ファイル | ライセンス | アクション |
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AHC30_401.pdf (6.4 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2010-08-06 | |||||
タイトル | ||||||
タイトル | A Novel Amplification Method of Nonradioactive In Situ Hybridization Signal for Specific RNA with Biotinylated Tyramine | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | In situ hybridization | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | 28S rRNA | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Oligonucleotide | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Biotinylated tyramine | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Rat brain | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者 |
Koji, Takehiko
× Koji, Takehiko× Kanemitsu, Yoshiro× Hoshino, Akiko× Nakane, Paul K. |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | For a better performance of nonradioactive in situ hybridization for specific RNA sequences, signal amplification is sometimes required especially in the use of clinical specimens processed under suboptimal conditions and in the analysis of gene expression with very low reiteration. In the present study, we examined the usefulness of catalyzed signal amplification (CSA) or catalyzed reporter deposition system with biotinylated tyramine to amplify colorimetric in situ hybridization signals. Our CSA protocol included the use of biotinylated tyramine after the reaction of thymine-thymine (T-T) dimerized DNA with horseradish peroxidase (HRP) labeled anti-(T-T dimer) antibody, followed by the reaction with HRP-labeled streptavidin. When thymine-thymine dimerized λ phage DNA was spotted onto a nitrocellulose filter at 1pg-10ng and then detected by the direct immunoperoxidase method, the indirect immunoperoxidase method or the CSA method, the CSA method gave the highest sensitivity with about 100-fold increase compared to that of the direct method. Next, in the frozen sections of rat brain, which was fixed with 4% paraformaldehyde, 28S rRNA staining by in situ hybridization with only 10ng/ml of T-T dimerized oligonucleotide probe complementary to rat 28S rRNA was compared among the direct method, the HRP-labeled avidinbiotin complex method and the CSA method. Again, intense 28S rRNA signal was obtained only with the CSA method. Therefore, we confirmed the usefulness of the catalyzed deposition system with biotinylated tyramine in nonradioactive colorimetric in situ hybridization for specific RNA sequences in tissue sections. | |||||
書誌情報 |
Acta Histochemica et Cytochemica 巻 30, 号 4, p. 401-406, 発行日 1997-08-01 |
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出版者 | ||||||
出版者 | 日本組織細胞化学会 | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 00445991 | |||||
EISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 13475800 | |||||
書誌レコードID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AA00508022 | |||||
権利 | ||||||
権利情報 | Copyright © 1997 By the Japan Society of Histochemistry and Cytochemistry | |||||
著者版フラグ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
引用 | ||||||
内容記述タイプ | Other | |||||
内容記述 | Acta Histochemica et Cytochemica, 30(4), pp.401-406; 1997 |