ThermoFisher (Life Tech)
Click-iT® EdU Pacific Blue™ Flow Cytometry Assay Kit
C10418
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This kit was used to monitor neutrophil production and maturation in vivo (murine) - it works great and is generally easy to use. The kit gives you plenty of reagents for many assays, the first thing I ran out of was EdU itself, and that can be easily purchased separately (and for much cheaper) so you can get full use of all the reagents.
EdU staining for flow cytometry
Murine blood and bone marrow
Inject EdU i.p. into mice, collect samples at various timepoints (for me, I was looking at neutrophil production, so I looked at 1, 2, 3, 4 days post injection). Peripheral blood or bone marrow was used. Collect cells, lysed red blood cells, surface stain and fix. Then follow EdU Click protocol as per guidelines. I scaled this down so I could do all my staining in a 96-well plate - this worked perfectly, kept all ratios of reagents the same, just did the staining in 200uL.
Can scale down reactions (as long as you don't have a ton of cells to stain) so it fits in a 96-well plate and that way you get more samples out of the kit. I ran out of EdU first because I was doing in vivo studies and had to inject more i.p. than you would use in tissue culture - but EdU is easy to buy separately - though I havent actually tried this yet, theres no reason it shouldn't be compatable.
Consistent, bright staining - analyzed by flow cytometry. On the left is a sample taken 24hr post injection of EdU (PMN don't proliferate, so they won't take up EdU, so they will only be EdU positive when the precursor stem cells proliferate and take up EdU, then mature and get released into the blood - so nothing is positive). The right shows a sample at 48 hrs, by which time new PMN have been made, and there is a very clear EdU positive population.
None
Easy to use
Expensive (similar to most flow products, but still, a consideration)
Reliable, will use again.
Product Name: Click-iT EdU Pacific Blue Flow Cytometry Assay KitSupplier Page