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. 2014 Sep 13;7:643. doi: 10.1186/1756-0500-7-643

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© Oh et al.; licensee BioMed Central Ltd. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Screening of primary transgenic plants expressing C-terminally FLAG-tagged HSP90.5. A, Schematic diagram of C-terminally FLAG-tagged HSP90.5 that was constructed in binary vector pGWB402Ω [41] through gateway cloning system. B, Examination of the specificity of polyclonal rabbit anti-HSP90.5 antibody. Immunoblotting was performed using purified cytosolic HSP90 from S. cerevisiae (yHSP82), Arabidopsis (AtHSP90.2) and human (hHSP90α), and ER-localized HSP90 from Arabidopsis (AtHSP90.7) and canine (dGRP94), with 50 ng loaded of each, and different amounts of purified AtHSP90.5. CBB staining represents SDS-PAGE stained with Coomassie brilliant blue. C, Immunoblotting of total leaf lysate proteins from 3-week-old primary transgenic seedlings. A total of 7.5 μg protein was separated by SDS-PAGE and immunoblotted with anti-FLAG, anti-HSP90.5 and anti-HSP90.2 antibodies.