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. 2009 Oct 22;9:254. doi: 10.1186/1471-2148-9-254

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Copyright © 2009 Him et al; licensee 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 cited.

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Hsp-90 from some clade III nematodes binds to GA. Extracts of various nematodes were analysed by immuno-blot (panels a and c) and by GA pull-down assay (panels b and d). Equivalent amounts of protein (determined by Bio-Rad protein assay) were analysed by SDS-PAGE on 10% gels and transferred to NCP. Blots were probed with 1:5000 rabbit anti-Brugia Hsp-90 antibody followed by 1:10,000 HRP-labelled anti-rabbit IgG. Bound antibody was detected by chemiluminesence. GA pull-down assays (panels b and d) were carried out with 500 μg of worm lysate using control or GA beads. Control beads were negative in all cases and are not shown. GA pull-down assays were analysed by SDS-PAGE and immuno-blotting as described above. Lane 1, B. pahangi, lane 2, A. viteae, lane 3 D. immitis, lane 4, O. ochengi, lane 5, B. pahangi, lane 6, T. cati, lane 7, A. suum, lane 8, P. equorum, lane 9, A. simplex, lane 10, P. decipens, lane 11, A. crassus.