BrdU proliferation assay during growth factor treatment - (Jul/11/2007 )

Hi,

I have a problem with BrdU assay (this is my first time).

I'm trying to do a proliferation assay on my cells in the presence of various growth factors. So I have to treat the cells with the factors for 2 days but at the same time I need to use BrdU to see how they proliferate.

According the the protocol, I have to add BrdU into the medium for 1 h and then aspirate the medium. Where exactly should my treatment go into this protocol? Should I remove BrdU from the environment before I do the treatment or is it necessary for BrdU to be present during all these treatments?

I'm short of time for the assay so the problem is urgent. I'll be very happy if someone shows me the light Thanks in advance

you add BrdU labelling solution, incubate for 1 hour and then without removing the labelling solution add your treatment medium. I don't know how long you are going to treat your cells but if it will be longer than generation time of your cells, you simply add BrdU as soon as the generation time is reached.
Hope it helps:)

It helps a lot I'm gonna try it. Thanks