C12FDG (solution)

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Solution preparation^[2]
1.1 Storage solution
Storage: After aliquoting, store at -20°C or -80°C in the dark and avoid repeated freezing and thawing.
1.2 Preparation of working solution
Dilute with PBS (or other buffer). The corresponding stock solution can be diluted according to the actual situation. Note that if the solvent is DMSO, the cytotoxicity of DMSO must be considered, and a solvent control should be prepared; if the solvent is pure water, the working solution needs to be filtered and sterilized before adding cells.
Note: The working solution should be prepared and used immediately, and protected from light.
2. Fluorescent senescence-associated β-galactosidase (SA-β-Gal) assay:
2.1 Culture cells in 6-, 12- , 24-, or 96-well plates at a density of 5× 10⁵ cells/mL overnight. Incubate the cells according to your normal protocol.
2.2 Wash cells by PBS once and fix with fixation solution (2% formaldehyde/0.2% glutaraldehyde in distilled water) at room temperature for 5 min.
2.3 Wash cells by PBS two times, and stain with 33 μM C12FDG for 10 min, and with 1 μg/mL Hoechst 33342 (HY-15559) for 10 min.
2.4 Image these cells by a 20× objective and 360-nm (Hoechst 33342) and 480-nm (C12FDG) excitation filters, and monitor through 460-nm and 535-nm emission filters, respectively.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.