Re: M-phase resolution
monard@adarc.nyu.edu
Thu, 15 Feb 1996 11:57:35 -0400
RE:
>Hello All,
>There is a researcher here interested in looking at expression of a
>nuclear antigen and how the expression varies relative to the cell
>cycle. We've done PI vs. FITC with great results, but we would like
>to better resolve the mitotic cells. What is the best method(s) for
>doing this?
>Thanks,
>Susan Lee
>Children's Hospital Medical Center
>Cincinnati, Ohio
>(513) 559-4769
>lees0@CHMCc.org
Hi
Maybe you could use colcemid to block your cells in mitosis then do your
PI and FITC. After 12 hours or so depending on how fast your cells grow the
majority of cells in the G2/M peak will be mitotic. With adherent cell
lines, blocking with colcemid then using mitotic shake-off yields a
suspension of almost exclusivly mitotic cells.
Have fun.
Simon Monard
Aaron Diamond Center
NYC