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Coomassie brilliant blue G250 色素結合法による高感度タンパク質定量法
https://obihiro.repo.nii.ac.jp/records/1853
https://obihiro.repo.nii.ac.jp/records/18533d6ec2e0-995c-4496-ae37-46618ec8e9bf
名前 / ファイル | ライセンス | アクション |
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i061122-1.pdf (557.8 kB)
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Item type | 紀要論文 / Departmental Bulletin Paper(1) | |||||
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公開日 | 2006-11-22 | |||||
タイトル | ||||||
タイトル | Coomassie brilliant blue G250 色素結合法による高感度タンパク質定量法 | |||||
言語 | ||||||
言語 | jpn | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | CBB 色素結合法 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | 色素結合法 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | 蛋白質定量法 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | CBB dye-binding assay | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | dye-binding assay | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | protein assay | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | CBB G250 | |||||
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 | departmental bulletin paper | |||||
その他(別言語等)のタイトル | ||||||
その他のタイトル | Coomassie brilliant blue G250 dye-binding microassay for protein | |||||
著者 |
井熊, 武志
× 井熊, 武志× 竹内, 宏治× 匂坂, 慶子× 高澤, 俊英 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Coomassie brilliant blue G-250 (CBB)を用いてタンパク質定量用高感度試薬の調製を試みた。まず、Bradford reagent (0.01%(w/v) CBB-4.75%(v/v)エタノール-8.5%(w/v)リン酸)を使用し、Bradford (1976)の高感度発色系(0.1mLタンパク質溶液、1mL Bradford reagent)によってBSA(0-9μg)を標準物質として用いてBradford法を検証した。Bradford reagent (Bradford 1976)は、調製直後から経日保存中にCBB沈殿を生じ、使用する度毎にろ過操作を必要とし、従ってその度に標準曲線作成実験を行わなければならず、実験操作上煩雑で、この欠点はBradford reagent に改良を加えても解決できなかった。そこで、保存中にCBB沈殿を生じず、安定なCBB reagentの調製を試み、充分使用に耐えるCBB reagentを開発した。CBB reagentの濃度は、0.025%(w/v) CBB-12.5%(v/v)メタノール-70.83%(w/v)リン酸である。CBBによるタンパク質の発色系は、0.8mLタンパク質溶液(0-9μg protein)、0.2mL CBB reagentで行った。 | |||||
抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | The high sensitive reagent using Coomassie brilliant blue G250 (CBB) for protein microassay has been prepared. At first, the high sensitive coloration system (0.1mL protein solution and 1mL Bradford reagent) was examined with Bradford reagent (0.01%(w/v) CBB-4.75%(v/v) ethanol-8.5%(w/v) phosphoric acid) according to Bradford (1976) using BSA(0-9μg) as a standard. The Bradford reagent produced CBB precipitate just after preparation and during storage, so filtration of it was needed before every use. Therefore, calibration of the Bradford CBB reagent due to change in CBB concentration should be carried out, which required time. These demerits had not been overcome by our modifications of the Bradford reagent. Thus we developed a stable, sufficiently available CBB reagent without CBB precipitate during storage. The CBB reagent is the 0.025%(w/v) CBB-12.5%(v/v) methanol-70.83%(w/v) phosphoric acid. The protein coloration with CBB was done at 0.8mL protein solution(0-9μg protein) and 0.2mL CBB reagent. | |||||
書誌情報 |
帯広畜産大学学術研究報告. 自然科学 en : Research bulletin of Obihiro University. Natural science 巻 23, 号 1, p. 18-26, 発行日 2002-12 |
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出版者 | ||||||
出版者 | 帯広畜産大学 | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 13476556 | |||||
書誌レコードID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AA11877530 | |||||
権利 | ||||||
権利情報 | 帯広畜産大学 | |||||
フォーマット | ||||||
内容記述タイプ | Other | |||||
内容記述 | application/pdf | |||||
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 |