Detection of Biotinylated Proteins in Polyacrylamide Gels Using an Avidin–Fluorescein Conjugate
作者： Michihiro Nakamura , Kouhei Tsumoto , Kazunori Ishimura , Izumi Kumagai
关键词:
摘要: Biotinylated proteins are widely used as a molecular tool in biotechnological applications. In this paper, we demonstrated that biotinylated after electrophoresis were detected directly gels using an avidin-fluorescein conjugate with fluorescence image analyzer. Upon analysis of the purified and chemically protein, sensitivity method was almost equal to silver staining. Chemically Escherichia coli cell surfaces could also be specifically our method. Furthermore, recombinant fused biotin acceptor domain enzymatically vivo lysate E. specifically. The specificity high, procedure is simple. Therefore, would benefit detection via gel various fields study avidin-biotin technology.
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sci-hub.se 参考文章(13)
Jörg Willuda, Susanne Kubetzko, Robert Waibel, P. August Schubiger, Uwe Zangemeister-Wittke, Andreas Plückthun, Tumor Targeting of Mono-, Di-, and Tetravalent Anti-p185HER-2 Miniantibodies Multimerized by Self-associating Peptides Journal of Biological Chemistry. ,vol. 276, pp. 14385- 14392 ,(2001) , 10.1074/JBC.M011669200
Edward A. Bayer, Meir Wilchek, The use of the avidin-biotin complex as a tool in molecular biology Methods of biochemical analysis. ,vol. 26, pp. 1- 45 ,(2006) , 10.1002/9780470110461.CH1
J E Cronan, Biotination of proteins in vivo. A post-translational modification to label, purify, and study proteins Journal of Biological Chemistry. ,vol. 265, pp. 10327- 10333 ,(1990) , 10.1016/S0021-9258(18)86949-6
Bhami C. Shenoy, Harland G. Wood, Purification and properties of the synthetase catalyzing the biotination of the aposubunit of transcarboxylase from Propionibacterium shermanii. The FASEB Journal. ,vol. 2, pp. 2396- 2401 ,(1988) , 10.1096/FASEBJ.2.8.3360240
Gunilla Høyer-Hansen, Maria J.A.G Hamers, Anders N Pedersen, Hans Jørgen Nielsen, Nils Brünner, Keld Danø, Ross W Stephens, Loss of ELISA specificity due to biotinylation of monoclonal antibodies Journal of Immunological Methods. ,vol. 235, pp. 91- 99 ,(2000) , 10.1016/S0022-1759(99)00222-7
Naoko Yamano, Yoshikazu Kawata, Hiroyuki Kojima, Koji Yoda, Makari Yamasaki, In vivo biotinylation of fusion proteins expressed in Escherichia coli with a sequence of Propionibacterium freudenreichii transcarboxylase 1.3S biotin subunit. Bioscience, Biotechnology, and Biochemistry. ,vol. 56, pp. 1017- 1026 ,(1992) , 10.1271/BBB.56.1017
Rüdiger Graf, Peter Friedl, Detection of Immobilized Proteins on Nitrocellulose Membranes Using a Biotinylation-Dependent System Analytical Biochemistry. ,vol. 273, pp. 291- 297 ,(1999) , 10.1006/ABIO.1999.4237
Walter L. Hurley, Eve Finkelstein, Brent D. Holst, Identification of surface proteins on bovine leukocytes by a biotin-avidin protein blotting technique Journal of Immunological Methods. ,vol. 85, pp. 195- 202 ,(1985) , 10.1016/0022-1759(85)90287-X
Annie-Paule Sibler, Emmanuelle Kempf, Arnaud Glacet, Georges Orfanoudakis, Dominique Bourel, Etienne Weiss, In vivo biotinylated recombinant antibodies: high efficiency of labelling and application to the cloning of active anti-human IgG1 Fab fragments. Journal of Immunological Methods. ,vol. 224, pp. 129- 140 ,(1999) , 10.1016/S0022-1759(99)00016-2
Meir Wilchek, Edward A. Bayer, Avidin-biotin technology ten years on: has it lived up to its expectations? Trends in Biochemical Sciences. ,vol. 14, pp. 408- 412 ,(1989) , 10.1016/0968-0004(89)90289-2