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cyclin B1 staining during prometaphase and metaphase

CyclinB1 cell cycle metaphase

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#1 dcandas

dcandas

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Posted 03 February 2014 - 11:25 AM

Hi there,

 

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.

 

Thanks







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