C12FDG (Synonyms: 5-Dodecanoylaminofluorescein di-β-D-Galactopyranoside)

Name: C12FDG
Brand: GlpBio
SKU: GC43021
Availability: InStock
Rating: 5 (35 reviews)

Catalog No.GC43021

C12FDG releases luciferin by enzymolysis of β-galactosidase, and the fluorescence of luciferin can be used to quantify the activity of β-galactosidase. The maximum excitation/emission of fluorescein is 485/530 nm.

Products are for research use only. Not for human use. We do not sell to patients.

C12FDG Chemical Structure

Cas No.: 138777-25-0

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500ug	$130.00	In stock
1mg	$200.00	In stock
5mg	$600.00	In stock

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Sample solution is provided at 25 µL, 10mM.

Product has been cited by 1 publications

Brain Behav Immun 103 (2022): 85-96.PMID:35427759

Description Protocol Chemical Properties Product Documents

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Quality Control & SDS

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Purity: >98.00%

Protocol

Protocol for Fluorescent aging associated β-galactosidase (SA-β-Gal) experiment using C12FDG^[1]:

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 protocol only provides a guideline, and should be modified according to your specific needs.

Protocol for Measurement of Senescence-Associated β-galactosidase activity using C12FDG^[2]:

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.

Endothelial cells (ECs) were alkalinized (pH 6) with chloroquine (300 μM) for 1h, followed by 1 h incubation with C12FDG (33 μM), a fluorogenic substrate of SA-β-gal .
Cells were thereafter washed with ice-cold PBS, harvested with trypsin and freshly analyzed using the CellQuest software. Light scattering parameters were set to eliminate dead cells and subcellular debris.
The green C12-fluorescein signal was measured and SA-β-gal activity estimated using the mean fluorescence intensity (MFI) of the cell population.

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.

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.

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	Soluble in DMSO	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.
Shipping Condition	Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.

Complete Stock Solution Preparation Table

Prepare stock solution
		1 mg	5 mg	10 mg
	1 mM	1.1711 mL	5.8555 mL	11.711 mL
	5 mM	0.2342 mL	1.1711 mL	2.3422 mL
	10 mM	0.1171 mL	0.5855 mL	1.1711 mL

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In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

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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.)

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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 ddH₂O, 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.

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Average Rating: 5 ★★★★★ (Based on Reviews and 35 reference(s) in Google Scholar.)

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C12FDG (Synonyms: 5-Dodecanoylaminofluorescein di-β-D-Galactopyranoside)

Customer Reviews

GlpBio Citations

Bioactive Compounds Premium Provider

Product has been cited by 1 publications

Product Documents

Quality Control & SDS

Protocol

Complete Stock Solution Preparation Table

Molarity Calculator

Dilution Calculator

In vivo Formulation Calculator (Clear solution)

Reviews

C12FDG (Synonyms: 5-Dodecanoylaminofluorescein di-β-D-Galactopyranoside)

Customer Reviews

GlpBio Citations

Bioactive Compounds Premium Provider

Product has been cited by 1 publications

Product Documents

Quality Control & SDS

Protocol

Complete Stock Solution Preparation Table

Molarity Calculator

Dilution Calculator

In vivo Formulation Calculator (Clear solution)

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Reviews