Fig. 7. Effect of XBub3 antibodies on XBub3 and XMad2 localisation in egg extracts. Metaphase chromosomes were assembled in egg extracts and treated with (+noc) or without (-noc) nocodazole. Sheep peptide or full-length XBub3 antibodies were added to the extracts 90 minutes before nocodazole addition. Chromosomes were isolated through a 30% glycerol cushion onto coverslips and stained with a sheep peptide XBub3 antibody followed by a fluorescent anti-sheep antibody (-noc, +noc) or fluorescent anti-sheep antibody alone (+noc +sheep XBub3 peptide Ab, +noc +sheep GST XBub3 Ab) to detect the sheep antibodies pre-incubated in the egg extract. Co-staining with XMad2 was achieved by incubating the coverslips with the rabbit XMad2 antibody. Chromosomes were detected by mounting the coverslips in medium containing DAPI (DNA).