Facs for BrdU and other staining
Posted 29 July 2010 - 08:04 AM
I am trying to use FACS to analyze the co-expression of GFP, BrdU and other transcription factor expression from mouse brain right now,
but our lab is not really experienced with FACS and I dont even know it works or not.
Especially, BrdU staining require denaturation with acid so I am wondering anyone have any comment about this???
We are also considering useing DNAse treatment instead of acid, but still not sure wheather we can keep detectable GFP expression.
Any suggestion help, so please let me know if you know the protocol or any papers you guys can suggest me to read.
Thanks in advance
Posted 29 July 2010 - 01:57 PM
Roche puts out a kit that claims to be advantageous by using nucleases rather than acid, base or heat to denature DNA but their protocol calls for a fixative with a pH of 2 so this is kinda bogus. Plus the kit never worked well for me.
Edited by rkay447, 29 July 2010 - 02:06 PM.
Posted 24 August 2010 - 08:33 AM
Thank you for the detail reply.
Since I do use cell from animal, so I probably need to modify your protocol a bit,
but I just optimize protocol for SOX2 and GFP, so hopefully incorporating your reply will give good result!!!
Thanks again, and sorry for the late reply