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In-gel protein phosphatase assay using fluorogenic substrates
https://kagawa-u.repo.nii.ac.jp/records/525
https://kagawa-u.repo.nii.ac.jp/records/525ca38e53e-3dc1-48ae-89ee-4a7b48fab196
名前 / ファイル | ライセンス | アクション |
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AA00524867_400_1_118 (664.1 kB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||||||||||||||
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公開日 | 2012-03-27 | |||||||||||||||||
タイトル | ||||||||||||||||||
言語 | en | |||||||||||||||||
タイトル | In-gel protein phosphatase assay using fluorogenic substrates | |||||||||||||||||
言語 | ||||||||||||||||||
言語 | eng | |||||||||||||||||
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 | |||||||||||||||||
アクセス権 | ||||||||||||||||||
アクセス権 | open access | |||||||||||||||||
アクセス権URI | http://purl.org/coar/access_right/c_abf2 | |||||||||||||||||
著者 |
亀下, 勇
× 亀下, 勇
WEKO
482
× 馬場, 裕美× 梅田, 嘉徳× 末吉, 紀行
WEKO
2790
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抄録 | ||||||||||||||||||
内容記述タイプ | Abstract | |||||||||||||||||
内容記述 | We developed a method for the detection of phosphatase activity using fluorogenic substrates after polyacrylamide gel electrophoresis. When phosphatases such as Ca2+/calmodulin-dependent protein kinase phosphatase (CaMKP), protein phosphatase 2C (PP2C), protein phosphatase 5 (PP5), and alkaline phosphatase were resolved by polyacrylamide gel electrophoresis in the absence of SDS and the gel was incubated with a fluorogenic substrate such as 4-methylumbelliferyl phosphate (MUP), all of these phosphatase activities could be detected in situ. Although 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP) as well as MUP could be used as a fluorogenic substrate for an in-gel assay, MUP exhibited lower background fluorescence. Using this procedure, several fluorescent bands that correspond to endogenous phosphatases were observed after electrophoresis of various crude samples. The in-gel phosphatase assay could also be used to detect protein phosphatases resolved by SDS-polyacrylamide gel electrophoresis. In this case, however, the denaturation/renaturation process of resolved proteins was necessary for the detection of phosphatase activity. This procedure could be used for detection of renaturable protein phosphatases such as CaMKP and some other phosphatases expressed in cell extracts. The present fluorescent in-gel phosphatase assay is very useful, since no radioactive compounds or no special apparatus are required. | |||||||||||||||||
言語 | en | |||||||||||||||||
書誌情報 |
en : Analytical biochemistry 巻 400, 号 1, p. 118-122, 発行日 2010-05-01 |
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出版者 | ||||||||||||||||||
言語 | en | |||||||||||||||||
出版者 | Elsevier | |||||||||||||||||
ISSN | ||||||||||||||||||
収録物識別子タイプ | PISSN | |||||||||||||||||
収録物識別子 | 1096-0309 | |||||||||||||||||
書誌レコードID | ||||||||||||||||||
収録物識別子タイプ | NCID | |||||||||||||||||
収録物識別子 | AA00524867 | |||||||||||||||||
論文ID(NAID) | ||||||||||||||||||
関連タイプ | isVersionOf | |||||||||||||||||
識別子タイプ | NAID | |||||||||||||||||
関連識別子 | 80020966117 | |||||||||||||||||
PubMed番号 | ||||||||||||||||||
識別子タイプ | PMID | |||||||||||||||||
関連識別子 | 20045670 | |||||||||||||||||
権利 | ||||||||||||||||||
言語 | en | |||||||||||||||||
権利情報 | Copyright © 2009 Elsevier Inc. All rights reserved. | |||||||||||||||||
関連サイト | ||||||||||||||||||
関連タイプ | isVersionOf | |||||||||||||||||
識別子タイプ | DOI | |||||||||||||||||
関連識別子 | https://doi.org/10.1016/j.ab.2009.12.035 | |||||||||||||||||
著者版フラグ | ||||||||||||||||||
出版タイプ | AM | |||||||||||||||||
出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||||||||||||||
キーワード | ||||||||||||||||||
言語 | en | |||||||||||||||||
主題Scheme | Other | |||||||||||||||||
主題 | Fluorogenic substrate | |||||||||||||||||
キーワード | ||||||||||||||||||
言語 | en | |||||||||||||||||
主題Scheme | Other | |||||||||||||||||
主題 | In-gel assay | |||||||||||||||||
キーワード | ||||||||||||||||||
言語 | en | |||||||||||||||||
主題Scheme | Other | |||||||||||||||||
主題 | Native-PAGE | |||||||||||||||||
キーワード | ||||||||||||||||||
言語 | en | |||||||||||||||||
主題Scheme | Other | |||||||||||||||||
主題 | Protein phosphatase | |||||||||||||||||
キーワード | ||||||||||||||||||
言語 | en | |||||||||||||||||
主題Scheme | Other | |||||||||||||||||
主題 | SDS–PAGE | |||||||||||||||||
KEID | ||||||||||||||||||
4979 |