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How to avoid the reduction of extracellular staining fluorescence in BrdU protoc


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#1 MinhQuan

MinhQuan

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Posted 09 September 2012 - 07:28 PM

Hi everyone
I am experiencing lots, lots of difficulties in BrdU staining for NKT cells. I have to use 6 colors (NK1.1 -PE Cy7, CD4-V500, CD44-V450, CD8-APC cy7, CD1d-tetramer PE, TCRb FITC for extracellular staining of NKT cells subsets and APC for intracellular BrdU staining. I have tried 2 fix/perm buffers - already have in my lab (one from Biolegend and the other from Santa Cruz).

However, after fix/perm procedure (follow these product's protocol), several fluorescences (like PerCP Cy5.5, PE Cy7, PE disappear. I want to buy the kit from BD but I don't know whether it will work with all colors I need or not.

Are there anyone using the kit from BD? Could you please give me some advice? I would like to say "thank you" in advance. I think any advice would be very helpful for me because I do not have much experience in this protocol





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