C12FDG

C12FDG (5-Dodecanoylaminofluorescein di-β-D-Galactopyranoside) is a lipophilic green fluorescent substrate for β-galactosidase detection. C12-FDG is more sensitive than FDG (HY-101895) for beta-galactosidase activity determinations in animal cells[1].

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Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). 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. Wash cells by 200 μL of PBS once and fix with 100 μL of fixation solution (2% formaldehyde/0.2% glutaraldehyde in distilled water) at room temperature for 5 min. 3. Wash cells by 200 μL of PBS two times, and stain with 100 μL of 33 μM C12FDG (in PBS, pH=6.0) for 10 min, and with 200 μL of Hoechst solution (1 μg/mL Hoechst 33342 (HY-15559) in PBS, pH 6.0) for 10 min. 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.

[1]. Plovins A, et al. Use of fluorescein-di-beta-D-galactopyranoside (FDG) and C12-FDG as substrates for beta-galactosidase detection by flow cytometry in animal, bacterial, and yeast cells. Appl Environ Microbiol. 1994 Dec;60(12):4638-41.

[2]. Udono M, et al. Quantitative analysis of cellular senescence phenotypes using an imaging cytometer. Methods. 2012 Mar;56(3):383-8.