FAO Blue (Fatty Acid Oxidation Detection Reagent)
FAOBlue Directly Detect Fatty Acid Oxidation (FAO) Activity in LIVE CELLS. FAO Blue is a newly designed fluorescent detection system (Excitation 405nm: Emission 460nm) to study catabolic pathway for energy production. Mitochondria and associated enzymes in the catabolic pathways can be detected to measure FAO activities by simply adding the dye to your cell.
Cat.No. FNK-FDV-0033
Size 0.2 mg
Storage -20 ºC
Formulation : C24H31NO9
Molecular weight : 447.51 g/mol
Solubility : Soluble in DMSO
Description
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Absorbance and fluorescence spectrum of FAOBlue and coumarin-derivative |
Absorbance (left) and fluorescence (right) spectra of FAOBlue (blue line) and the coumarin-derivative dye released after metabolization of FAO (red line) in PBS buffer (pH 7.4).
Absorbance spectra: an absorption peak of coumarin-derivative dye is clearly shifted by FAO from the peak of FAOBlue.
Fluorescence spectra: Coumarin-derivative dye shows strong blue fluorescence but FAOBlue emits little fluorescence, when both compounds are exited at 405 nm. NOTE: FAOBlue shows blue fluorescence (gray line; 370-450 nm) when it is excited at 300-380 nm (max 350 nm). |
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Experimental guide for imaging |
Confocal laser microscopy: Please use 405 nm laser equipped in the microscopy. Using 405 nm laser allows to detect only a fluorescent signal from coumarin-derivative dye.
Epifluorescence microscopy: Excitation filter is very important. Commercial DAPI filters are not compatible with this reagent, because DAPI filters excite both FAOBlue and coumarin-derivative dye. Excitation filters which pass 390-450 nm wavelength light are recommended. |
Visualization of FAO activities in 4 cancer cell lines | |
Four cancer cell lines (HepG2, LNCaP, HeLa and A549) were treated with FAOBlue in HBS buffer with or without pre-treatment of etomoxir (40 uM, 3 hours), a potent FAO inhibitor. After FAOBlue incubation, blue fluorescence (Ex. 405 nm/ Em. 430-480 nm) was observed. All cell lines showed blue fluorescence in cytosol, but pre-treatment of etomoxir clearly decreased fluorescent intensities. These results indicated the blue fluorescence was derived from FAO activity in the cells.
* Experimental condition:
HepG2, 5 uM FAOBlue for 30 min.
LNCaP, 20 uM FAOBlue for 120 min HeLa, 20 ?M FAOBlue for 120 min A549, 5 ?M FAOBlue for 30 min |
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Perturbation of FAO activity by drugs | |
HepG2 cells were pre-incubated with 200 uM AICAR, a FAO activator via AMPK activation, for 3 hours or 200 uM of ranolazine, a partial FAO inhibitor, for 12 hours. After drug treatment, the cells were incubated with 5 uM FAOBlue for 30 min. Compared with control cells, pretreatment with AICAR significantly increased blue fluorescent intensity. On the other hand, pretreatment with the partial FAO inhibitor ranolazine clearly decreased blue fluorescent intensity. Ex. 405 nm/ Em. 430-480 nm
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Quantitative analysis of the drug effects on FAO activity | |
ND630 is an inhibitor of acetyl-CoA carboxylase and considered as a potential therapeutic drug for non?alcoholic fatty liver disease (NAFLD). HepG2 cells were pre-incubated with various concentration of ND630 for 4 hours. After ND630 treatment, cells were treated with 5 uM FAOBlue for 30 min. Blue fluorescent intensities of each concentration of ND630 were quantified. Ex. 405 nm/ Em. 430-480 nm
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Analysis of FAO activity using NASH model mouse | |
Non-alcoholic steatohepatitis (NASH) is a typical disease which shows low metabolic activity of FAs. Control healthy mice and NASH model mice were orally administered with 400 mg/kg bezafibrate, a therapeutic agent of NASH. After 4 weeks administration, primary hepatocytes were isolated from control mice and NASH model mice and cultured in culture dishes. Primary hepatocytes were further treated with 5 uM FAOBlue for 30 min and fluorescence imaging was performed. Compared with control cells, NASH model mouse-derived hepatocytes showed low FAO activity. Bezafibrate dramatically recovered FAO activity of hepatocytes isolated from NASH model mouse. FAOBlue is a powerful tool to estimate drug effects and efficiency on FAO activity. *Detail procedure of mice experiment is described in Ref.1. Ex. 405 nm/ Em. 430-480 nm |
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We are pleased to announce that the product article about FAOBlue was posted on BioTechniques which is one of the famous international journal regarding life sciences since September 1. |