C12FDG (solution) 

C12FDG (5-Dodecanoylaminofluorescein di-β-D-Galactopyranoside) (solution) is a lipophilic green fluorescent substrate for β-galactosidase detection. C12-FDG is more sensitive than Fluorescein di(β-D-galactopyranoside) (HY-101895) for beta-galactosidase activity determinations in animal cells (Ex/Em = 488/523 nm)^[1].
Solution Concentration: 10 mM

Guidelines (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 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.
2.3 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.
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.

853.92

C₄₄H₅₅NO₁₆

138777-25-0

CCCCCCCCCCCC(NC1=CC2=C(C3(C4=C(OC5=C3C=CC(O[C@H]6[C@@H]([C@H]([C@H]([C@@H](CO)O6)O)O)O)=C5)C=C(O[C@H]7[C@@H]([C@H]([C@H]([C@@H](CO)O7)O)O)O)C=C4)OC2=O)C=C1)=O

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [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.  [Content Brief]

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