FITC-Palmitic acid

1.PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 C after preparation. Avoid repeated freeze-thaw cycles

FITC-Palmitic acid stock solution

Prepare a 5 to 10 mM stock solution by dissolving an appropriate amount of FITC-Palmitic acid in DMSO.

Note: For example, to prepare a 5 mM DMSO stock solution dissolve1 mg of the FITC-Palmitic acid in 235 uL of DMSO.

2.PREPARATION OF WORKING SOLUTION

FITC-Palmitic acid working solution

Prepare a 5 to 10 μM FITC-Palmitic acid working solution by diluting the stock solution in HHBS.

Note: Prepare a fresh working solution before use

3.SAMPLE EXPERIMENTAL PROTOCOL

The following protocol can be used as a guideline. lf needed, optimize the protocol to achieve the desired results.

1. Culture cells as desired.

2. Remove cell culture medium.

Note: Cell culture medium must be removed since it may interfere with the conjugate staining.

3. Add 100 uL of FITC-Palmitic acid working solution per well.

4. Incubate the cells in the dark for 30 minutes at roomtemperature.

5. Remove the FITC-Palmitic acid working solution and wash cells twice with HHBS.

6. Add HHBS buffer to the cells and observe the cells using a fluorescence microscope with a FITC filter set.

7. Optional: The cells can be fixed after staining.

Image of Hela cells stained with FITC-Palmitic acid and then fixed with 4% formaldehyde. Live Hela cells were cultured in a 96-well plate overnight, and the medium was removed before staining. 100 µL of the FITC-Palmitic acid (10 µM) in HHBS buffer was added to each well. Cells were stained for 30 minutes at room temperature, washed twice with HHBS buffer, and then fixed with 4% formaldehyde. The cells were imaged with a FITC filter before and after fixation.