Interaction of a novel biotin-tagged photoaffinity probe with its target protein, VEGFR-2

Bulletin of the Korean Chemical Society

Volumn 27, Issue 9, 2006, Pages 1285-1286

  Myung, Hee Kim ^a   Han, Sun Young ^a,b   Bum, Tae Kim ^a   Yong, Ki Min ^a   Seong, Hwan Kim ^a  

^a Korea Research Institute of Chemical Technology   (South Korea)

^b UNIVERSITY OF TEXAS AT DALLAS   (United States)

Author keywords

Biotin tagged photoaffinity probe; Ligand protein interaction; Vascular endothelial growth factor receptor 2; VEGFR 2

Indexed keywords

EID: 33749030457     PISSN: 02532964     EISSN: 12295949     Source Type: Journal    
DOI: 10.5012/bkcs.2006.27.9.1285     Document Type: Article

References (16)

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14. 16 Electrophoresis was performed using the Mini Protean 3 Cell (Bio-Rad). Proteins separated on the gels were transferred onto nitrocellulose membrane (Scheicher & Schnell BioScience, Germany), and the membrane was incubated in blocking buffer (10 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% Tween 20, 3% nonfat dry milk). The membrane was incubated for 2 hr at room temperature with 1:1000 diluted primary antibodies. Horse radish peroxidase(HRP)-conjugated anti-VEGFR-2 was purchased from Santa Cruz Biotechnology Inc. (CA). HRP-conjugated streptavidin was purchased from Merck and the goat anti-biotin purchased from Pierce. After washing three times for 15 min with blocking buffer, and developed with SuperSignal West Femto Maximum Sensitivity Substrate (Pierce). When used the goat anti-biotin, membrane was incubated for 1 hr at room temperature with 1:2000 diluted HRP labeled anti-goat IgG (Pierce) before the development. The immunoreactivity was detected using LAS-3000 luminescent image analyzer (Fuji Photo Film Co., Ltd., Japan).
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