@article{oai:obihiro.repo.nii.ac.jp:00001853,
 author = {Ikuma, Takeshi and 井熊, 武志 and 竹内, 宏治 and 匂坂, 慶子 and 高澤, 俊英},
 issue = {1},
 journal = {帯広畜産大学学術研究報告. 自然科学, Research bulletin of Obihiro University. Natural science},
 month = {Dec},
 note = {application/pdf, 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。, 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.},
 pages = {18--26},
 title = {Coomassie brilliant blue G250 色素結合法による高感度タンパク質定量法},
 volume = {23},
 year = {2002}
}