BrdU by immunofluorescence. Help! - (Feb/15/2011 )
I'm trying to do a BrdU proliferation assay for some glial precursor cells by immunofluorescence. My problem is that I need to stain for markers other than BrdU in order to identify my cells as it is not a 100% pure culture. At the moment I'm doing it by running start to finish my normal protocol for my marker (and DAPI) and then through a protocol for BrdU staining, with a HCl step to denature. But at the end, all the cells look a bit of a mess and my BrdU is very faint. I'm using 4%PFA to fix first and then methanol to fix before BrdU staining.
If anyone has a protocol that works I'd really appreciate it!
You could try Click-iT from Invitrogen http://tools.invitrogen.com/content.cfm?pageid=12145&cid=fl-EDU, it worked for me quite ok.
BrdU is always a big mess...
Rsm on Thu Feb 17 12:42:13 2011 said:
You could try Click-iT from Invitrogen http://tools.invitrogen.com/content.cfm?pageid=12145&cid=fl-EDU, it worked for me quite ok.
BrdU is always a big mess...
Thanks very much for that! I'm going to try and get a sample of it, sounds good. Glad to hear I'm not the only one who finds BrdU to be messy
