C12FDG (5-Dodecanoylaminofluorescein di-β-D-Galactopyranoside) 是用于 β-半乳糖苷酶检测的亲脂性绿色荧光底物。C12-FDG 比 FDG (HY-101895) 对动物细胞 β-半乳糖苷酶的活性测定更敏感。

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.

Solid

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:
1. Culture cells in 6-, 12- , 24-, or 96-well plates at a density of 5× 10 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

Room temperature in continental US; may vary elsewhere.

4°C, protect from light In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

[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.