Phos-tag beads as an immunoblotting enhancer for selective detection of phosphoproteins in cell lysates
Analytical Biochemistry 389 巻 1 号
83-85 頁
2009-06-01 発行
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| ファイル情報(添付) | |
| タイトル ( eng ) |
Phos-tag beads as an immunoblotting enhancer for selective detection of phosphoproteins in cell lysates
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| 作成者 |
Kinoshita-Kikuta Emiko
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| 収録物名 |
Analytical Biochemistry
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| 巻 | 389 |
| 号 | 1 |
| 開始ページ | 83 |
| 終了ページ | 85 |
| 抄録 |
The low specificity of anti-phosphoprotein antibodies is often a problem in immunoblotting analyses. We introduce a simple pretreatment procedure for cell lysates to give more-specific detection of phosphoproteins in immunoblotting. Cellular phosphoproteins were preferentially trapped on Phos-tag agarose phosphate-affinity beads in a homemade spin-centrifuge microtube unit, and nonphosphorylated proteins were excluded in the filtrate. The phosphoprotein-bound beads suspended in a sample-loading dye solution were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by Western blotting. We demonstrated improved detection of phosphorylated Shc and mitogen-activated protein kinase isoforms in A431 cell lysates by this new technique
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| 著者キーワード |
Phos-tag
Immunoblotting enhancer
Anti-phosphoprotein antibody
Phosphoprotein
Phosphorylation
Signal transduction
Western blotting
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| NDC分類 |
生物科学・一般生物学 [ 460 ]
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| 言語 |
英語
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| 資源タイプ | 学術雑誌論文 |
| 出版者 |
Elsevier
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| 発行日 | 2009-06-01 |
| 権利情報 |
Copyright (c) 2009 Elsevier Inc.
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| 出版タイプ | Author’s Original（十分な品質であるとして、著者から正式な査読に提出される版） |
| アクセス権 | オープンアクセス |
| 収録物識別子 |
[ISSN] 0003-2697
[ISSN] 1096-0309
[DOI] 10.1016/j.ab.2009.03.026
[NCID] AA00524867
[NCID] AA11537838
[DOI] http://dx.doi.org/10.1016/j.ab.2009.03.026
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