Poloxamer 407  (Cat.#:AYB26683)

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Dissolve 1 g of Poloxamer 407 in 10 mL distilled water to make a 10% (w/v) stock solution, or 2 g of Poloxamer 407 in 10 mL DMSO to make a 20% (w/v) stock solution. These may require heating from 40 to 50°C for about 30 minutes. Store solution at room temperature. Do not refrigerate or freeze the Poloxamer 407 solution since it may precipitate. If precipitation is observed, the precipitates can be dissolved by heating to 37°C and vortexing before use.
2. Dilute the 10% or 20% Poloxamer 407 stock solution into the cell-loading buffer such as Hanks and 20 mM Hepes buffer (HHBS) at 1:1000 to 1:500 dilution to achieve a 0.02 to 0.04% working solution.
3. The DMSO stock solution of AM ester is then diluted into the 0.02 to 0.04% working solution to achieve a final AM ester concentration between 1 μM and 10 μM. The final concentration of Poloxamer 407 is normally kept at or below 0.08%.
4. The cells are incubated at a desired temperature for between 10 minutes and 1 hour. In general it is desirable to use the minimum amount of AM ester needed to achieve adequate fluorescence signal to noise levels.
5. After labeling, the cells are washed with HHBS or fresh medium before starting the experiment.

The accuracy of these methods have not been independently confirmed. They are for reference only.