EdU is an alternative to BrdU from Life Technologies. On paper, it should work better than BrdU (of course). If anyone has experience with this method, could you share it with me? I'm particularly interested in labeling T-cells.
Hello Namjiin,
the EDU Kit is much easier to handle than the usual BrdU protocols and data look as good as you would like to see it from the BrdU experiments. There is not that much you can do wrong, when you follow the protocols from the companies. I attach a protocol from the FACS Laboratory, London Research Institute (credit goes to Derek Davies), which is quite detailed.
BUT
The click chemistry seems to interfere with the PE fluorochromes and their tandem conjugates, which you normally use for surface staining on immune cells (See troubleshooting in Dereks protocol). Also with quantum dots. So you cant do staining for EDU AFTER immunophenotyping using PE conjugates.
And just a last remark, the click chemistry is not that new, so there are other companies out there, that also sell the EDU kits for cell proliferation.
http://shop.bioanalytical-solutions.com/en/Click-Chemistry-based-products
Kind regards
Elmar
Hello Namjiin,
the EDU Kit is much easier to handle than the usual BrdU protocols and data look as good as you would like to see it from the BrdU experiments. There is not that much you can do wrong, when you follow the protocols from the companies. I attach a protocol from the FACS Laboratory, London Research Institute (credit goes to Derek Davies), which is quite detailed.
BUT
The click chemistry seems to interfere with the PE fluorochromes and their tandem conjugates, which you normally use for surface staining on immune cells (See troubleshooting in Dereks protocol). Also with quantum dots. So you cant do staining for EDU AFTER immunophenotyping using PE conjugates.
And just a last remark, the click chemistry is not that new, so there are other companies out there, that also sell the EDU kits for cell proliferation.
http://shop.bioanalytical-solutions.com/en/Click-Chemistry-based-products
Kind regards
Elmar
"On paper, it should work better than BrdU (of course)"
It does. The signal-to-noise ratio is much better, probably because the azide binds to less things than antibodies in cells. I had two logs between negative and positive cells, as compared to one log with BrdU.
- Badges
- Science method