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BrdU by immunofluorescence. Help!


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#1 LittleMissTcell

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Posted 15 February 2011 - 10:34 AM

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!

#2 Rsm

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Posted 17 February 2011 - 04:42 AM

You could try Click-iT from Invitrogen http://tools.invitro...2145&cid=fl-EDU, it worked for me quite ok.
BrdU is always a big mess...
I got soul, but I'm not a soldier

#3 LittleMissTcell

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Posted 21 February 2011 - 08:23 AM

You could try Click-iT from Invitrogen http://tools.invitro...2145&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 :)




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