Click-iT® Plus EdU Imaging KitsClick-iT® Plus EdU Imaging Kits kdorfman Mon, 08/12/2019 - 18:55
Materials required but not in kit:
|A (EdU)||10 mM||2 mL DMSO (C) or buffer|
|B (Alexa Fluor® picolyl azide)|
|D (reaction buffer)||10X|
|E Cu protectant|
|F (buffer additive)|
|G (Hoechst® 33342)|
Per tube, need 200uL
Buffer AdditiveBuffer Additive kdorfman Thu, 03/26/2020 - 14:18
To make a 10X stock solution of the Click-iT® EdU buffer additive (Component F): Add 2 mL deionized water to the vial, then mix until fully dissolved. After use, store any remaining stock solution at ≤–20˚C.
When stored as directed, this stock solution is stable for up to 1 year. If the solution develops a brown color, it has degraded and should be discarded.
EdUEdU kdorfman Wed, 09/11/2019 - 17:00
33mM (10X) Stock solution
100 mg of EdU powder (purchased from Carbosynth) https://www.carbosynth.com/carbosynth/website.nsf/(w-productdisplay)/D1…
is dissolved in
1.2 mL DMSO, then brought to 12 mL with H2O
Original recipe says: 10.8mL 0.5x E2 medium and
Rolf says the final concentration of DMSO should be more like 0.1% (after dilution of the 10X EdU to 1X in fish water)
Says to mix with water and warm gently (to thaw, use 60C for 15 minutes)
So try 33 mM EdU in 1% DMSO.
To make 20 mL 10X EdU in 1% DMSO, mix
- 166 mg EdU
- 0.2 mL DMSO (heat gently by putting it in a beaker of heated water)
- bring to final volume with distilled water
Reaction BufferReaction Buffer kdorfman Thu, 03/26/2020 - 14:13
Prepare a working solution of 1X Click-iT® EdU reaction buffer (Component D):
Transfer the solution (4 mL) in the Component D bottle to 36 mL of deionized water.
To make smaller amounts of 1X Click-iT® EdU reaction buffer,
dilute volumes from the Component D bottle 1:10 with deionized water.
After use, store any remaining 1X solution at 2–8˚C.
When stored as directed, this 1X solution is stable for 6 months.