Hi everyone,
For my neurogenesis project, I'd like to inject young pups (P14) with EdU for a long-term chase (P56). So far, my idea was to perform two days of pulse with 2 injections per day (P14 and P15) with a dose of 100 mg/kg per injection. Given how small the pups are at that time, it roughly corresponds to 50 - 60 µL of a solution of 10 mg/mL EdU. However, I'd like to know how to prepare such a solution. I've read protocols where EdU is first dissolved at 200 mg/mL in DMSO, then diluted in 1X PBS to 10 mg/mL. I am however worried that a 5% DMSO solution might be toxic for young pups. Has anyone ever injected young pups with EdU? Thanks for your help and answers!
https://assets.thermofisher.com/TFS-Assets/LSG/manuals/mp10044.pdf
You can prepare stock solution directly with PBS or other aqueous solution that you think is with less toxicity. You don't necessarily need to use DMSO.
I do embryonic, perinatal (P2) and postnatal (P21) EdU injection. My stock solution was prepared with PBS. I usually filter it to make sure it's sterile. For injection, I usually do 50mg/kg. For younger pups, you would have to dilute the stock solution further so the smallest needle (1ml syringe) can actually pick up enough solution (~50ul is a good start) for injection.
Hope this helps!
Thanks for your answer! What's the concentration of your stock solution? They preconize to do a 10 mM solution in DMSO or aqueous buffer (e.g. PBS) but if I do so, I'll have to inject 120 µL per mouse to obtain 50 µg/g bw (at P12 my mice are about 6 g).