Click-iT® Plus EdU Imaging Kits

Click-iT® Plus EdU Imaging Kits kdorfman Mon, 08/12/2019 - 18:55

Materials required but not in kit:

Stock Solutions

Component conc dilute with
A (EdU) 10 mM 2 mL DMSO (C) or buffer
B (Alexa Fluor® picolyl azide)
C (DMSO)
D (reaction buffer) 10X
E Cu protectant
F (buffer additive)
G (Hoechst® 33342)

Per tube, need 200uL

To make mL Click-iT cocktail
add: uL D (reaction buffer)
add: uL Copper protectant (E)
add: uL Alexa Fluor (B)
and: uL F (buffer additive)

Click-iT® Plus EdU Imaging Kits manual.pdf

Buffer Additive

Buffer 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.

EdU

EdU 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

1.2ml DMSO

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 Buffer

Reaction 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.