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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 clade V nematodes does not bind GA in a solid phase pull-down assay. Extracts of various nematodes were analysed by immuno-blots (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, C. elegans N2, lane 3, C. elegans daf-21 mutant, lane 4, O. tipulae, lane 5, P. pacificus, lane 6, B. pahangi, lane 7, T. circumcincta, lane 8, N. brasiliensis, lane 9, H. polygyrus, lane 10, H. contortus.