I am not familiar with cell cycle analysis and I was asked to perform a pro-metaphase and metaphase staining of MCF10A breast epithelial cells for CyclinB1, to see if the CyclinB1 antibody that I am using is specific and gives the known/usual pattern of CyclinB1 staining during these cell cycle phases.
Is there a chemical that will arrest the cells at pro-metaphase or metaphase? I have come across nocodazole but as far as I understand it arrests the cells by depolymerizing the microtubules. This doesn't work for me because I need intact mitotic spindle. CyclinB1 localizes to mitotic spindle during the metaphase and that's what I am supposed to show with my staining.
any advice is greatly appreciated.