C12FDG (Synonyms: 5-Dodecanoylaminofluorescein di-β-D-Galactopyranoside) |
Catalog No.GC43021
One-Click Copy Product Info
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C12FDG is the fluorescent substrate of β-galactosidase.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 138777-25-0
Sample solution is provided at 25 µL, 10mM.
C12FDG is the fluorescent substrate of β-galactosidase. By enzymolysis of β-galactosidase, fluorescein is released and its fluorescence can be used to quantify the activity of β-galactosidase. The excitation/emission maxima of fluorescein are 485/530 nm[1-3].
C12FDG(15 μM ; 2 h) staining can assess SA-β-Gal activity to detect senescence in THP-1 cells(15 μM ; 2 h) [4]and A172 cells(90 min; 33 µM) [5]. Live cell measurements of senescence-associated β-galactosidase (SA-β-Gal) activity using the fluorescent substrate C12FDG in combination with the determination of the total cell number using a DNA intercalating Hoechst dye opens the possibility to screen for senotherapeutic drugs [6].
References:
[1]. Zhang YZ, Naleway JJ, et,al. Detecting lacZ gene expression in living cells with new lipophilic, fluorogenic beta-galactosidase substrates. FASEB J. 1991 Dec;5(15):3108-13. doi: 10.1096/fasebj.5.15.1720751. PMID: 1720751.
[2]. Plovins A, Alvarez AM, 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. doi: 10.1128/aem.60.12.4638-4641.1994. PMID: 7811104; PMCID: PMC202038.
[3]. Fuhrmann-Stroissnigg H, Santiago FE, et,al. SA-β-Galactosidase-Based Screening Assay for the Identification of Senotherapeutic Drugs. J Vis Exp. 2019 Jun 28;(148). doi: 10.3791/58133. PMID: 31305507.
[4]. Díaz M, Pibuel M, et,al. 4-Methylumbelliferone induces antitumor effects independently of hyaluronan synthesis inhibition in human acute leukemia cell lines. Life Sci. 2021 Dec 15;287:120065. doi: 10.1016/j.lfs.2021.120065. Epub 2021 Oct 19. PMID: 34678263.
[5]. Beltzig L, Frumkina A, et,al. Cytotoxic, Genotoxic and Senolytic Potential of Native and Micellar Curcumin. Nutrients. 2021 Jul 13;13(7):2385. doi: 10.3390/nu13072385. PMID: 34371895; PMCID: PMC8308652.
[6].Waltz RA, Whitney KE, et,al. A Systemic and Local Comparison of Senescence in an Acute Anterior Cruciate Ligament Injury-A Pilot Case Series. Life (Basel). 2023 Jul 15;13(7):1567. doi: 10.3390/life13071567. PMID: 37511942; PMCID: PMC10381817.
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Protocol for Fluorescent aging associated β-galactosidase (SA-β-Gal) experiment using C12FDG [1]: |
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1. Cells were cultured with a density of 105 cells/mL. 2. The cells were washed once with 200 μL PBS, and then fixed with 100 μL fixing solution (2% formaldehyde /0.2% glutaraldehyde distilled water) at room temperature for 5 minutes. 3. The cells were washed with 200 μL PBS twice, stained with 100 μL 33 μM C12FDG (PBS, pH=6.0) for 10 min, and then 200 μL Hoechst solution (1 μg/mL Hoechst 33342) on PBS (pH 6.0) for 10 minutes. 4. These cells were imaged with a 20× objective lens and excitation filters of 360 nm (Hoechst 33342) and 480 nm (C12FDG), and monitored with emission filters of 460 nm and 535 nm, respectively. This C12FDG protocol only provides a guideline, and should be modified according to your specific needs. |
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Protocol for Measurement of Senescence-Associated β-galactosidase activity using C12FDG [2]: |
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The fluorogenic cell permeable substrate C12FDG was used to determine the senescence-associated β-galactosidase activity (SA-β-gal) in treated Endothelial cells by flow cytometry.
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References: [1]. Udono M, Kadooka K, et,al. Quantitative analysis of cellular senescence phenotypes using an imaging cytometer. Methods. 2012 Mar;56(3):383-8. doi: 10.1016/j.ymeth.2012.02.012. Epub 2012 Mar 3. PMID: 22406489. [2]. Qureshi AW, Altamimy R, et,al. Ageing enhances the shedding of splenocyte microvesicles with endothelial pro-senescent effect that is prevented by a short-term intake of omega-3 PUFA EPA:DHA 6:1. Biochem Pharmacol. 2020 Mar;173:113734. doi: 10.1016/j.bcp.2019.113734. Epub 2019 Dec 5. PMID: 31811867. |
| Cas No. | 138777-25-0 | SDF | |
| Synonyms | 5-Dodecanoylaminofluorescein di-β-D-Galactopyranoside | ||
| Chemical Name | N-[3',6'-bis(β-D-galactopyranosyloxy)-3-oxospiro[isobenzofuran-1(3H),9'-[9H]xanthen]-5-yl]-dodecanamide | ||
| Canonical SMILES | O[C@@H]([C@@H](O)[C@@H](CO)O1)[C@@H](O)[C@@H]1OC2=CC=C(C3(C4=CC=C(NC(CCCCCCCCCCC)=O)C=C4C(O3)=O)C(C=CC(O[C@@H]5O[C@H](CO)[C@H](O)[C@H](O)[C@H]5O)=C6)=C6O7)C7=C2 | ||
| Formula | C44H55NO16 | M.Wt | 853.9 |
| Solubility | DMSO : 50 mg/mL (58.55 mM; Need ultrasonic) | Storage | Store at -20°C, protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.1711 mL | 5.8555 mL | 11.711 mL |
| 5 mM | 234.2 μL | 1.1711 mL | 2.3422 mL |
| 10 mM | 117.1 μL | 585.5 μL | 1.1711 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 35 reference(s) in Google Scholar.)