cis-5,8,11,14,17-Eicosapentaenoic acid (EPA), metformin, compound C (AMPK inhibitor), insulin, STO-609 (CaMKK inhibitor), SB203580 (p38 MAPK inhibitor), fluo-3, AM and a monoclonal anti-β-actin antibody were purchased from Sigma Chemical Company (St. Louis, MO, USA). BAPTA-AM (intracellular calcium chelator), monoclonal antibody against ACC and polyclonal antibodies against GLUT4 were purchased from Abcam (Cambridge, MA, USA). Monoclonal antibodies against phosphorylated AMPKα, phosphorylated p38 MAPK, p38 MAPK, phosphorylated AS160, AS160, and the polyclonal antibody against AMPKα were obtained from Cell Signaling Technology (Danvers, MA, USA). The polyclonal antibody against phosphorylated ACC was provided by Merck (Rahway, NJ, USA). Fetal bovine serum (FBS) and penicillin-streptomycin were acquired from Thermo Fisher Scientific (Foster City, CA, USA). The monoclonal antibody against c-Myc was acquired from Santa Cruz Biotechnology (Dallas, TX, USA).
Fluo 3
Manufactured by Merck Group
Sourced in United States
Fluo-3 is a fluorescent calcium indicator used for the detection and measurement of calcium levels in biological samples. It functions by binding to calcium ions, which causes an increase in its fluorescence intensity. Fluo-3 is commonly used in various research applications, such as cell signaling studies, neuroscience, and drug discovery.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Anti β actin monoclonal antibody, by Merck Group (1 mentions)
Sb203580, by Merck Group (1 mentions)
Sto 609, by Merck Group (1 mentions)
Cis 5 8 11 14 17 eicosapentaenoic acid, by Merck Group (1 mentions)
Monoclonal antibody against acc, by Abcam (1 mentions)
Compound c, by Merck Group (1 mentions)
Metformin, by Merck Group (1 mentions)
Bapta am, by Abcam (1 mentions)
Filtermax f5 spectrophotometer, by Molecular Devices (1 mentions)
Leica tcs sp, by Leica camera (1 mentions)
Rhodamine b, by Merck Group (1 mentions)
Wortmannin, by Merck Group (1 mentions)
Von kossa staining kit, by Genmed Scientifics (1 mentions)
Rhodamine phalloidin, by Merck Group (1 mentions)
β glycerophosphate, by Merck Group (1 mentions)
6 protocols using fluo 3
1
AMPK and Calcium Signaling Modulators
2
Ca2+ Uptake Assay in Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Vesicles (2–400 μg protein) were added to 100 μl ice-cold Ca2+ uptake buffer (10 mM HEPES, 50 mM KCl, 2 mM MgCl2, 3% sucrose) containing 100 nM NaN3, 6.6 μM Fluo-3 (Sigma Aldrich, UK), 0.4 mM Mg2+-ATP and 12 μg/ml oligomycin. Free ionized Ca2+ was measured via Fluo-3 (490 nm excitation and 535 nm emission) in a 96-well plate, using a fluorometer (NovoStar).
3
Quantitative Calcium Flux Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Calcium assays were performed using Fluo-3 (Sigma Aldrich) solubilized to 10 mg/ml with dimethyl sulfoxide. Cells were loaded with 4 μg/ml Fluo-3 for 30 minutes at 37° C. Cells were washed three times in HEPES buffered saline solution (HBSS with 1 mM CaCl2, 0.5 mM MgCl2, 0.1% BSA, 10 mM HEPES) and resuspended to 1 × 106 cells/ml in HEPES buffered saline solution. For measurement of store-operated Ca2+ influx, 3 mM EGTA and 2 mM thapsigargin were added to deplete internal calcium stores. Ca2+ influx was induced by subsequent addition of 2 mM Ca2+ (free) after store depletion [24 (link), 39 (link)]. The emission wavelength set at (λem) 485 nm and capture excitation wavelength set (λex) at 520 nm. The relative fluorescence detection was performed using FilterMax F5 spectrophotometer (Molecular Devices, Sunnyvale, CA) and data obtained using instrument software.
4
Confocal microscopy analysis of sealers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the confocal laser scanning fluorescence microscopy analysis, the following dyes, respectively, were used at the time of filling: Fluo-3 (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany) for sealers based on bioceramic compounds (AH Plus Bioceramic, Bio-C Sealer and Bio-C Sealer Ion + ) and Rhodamine B (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany) for epoxy resin-based sealer (AH Plus). The 16 samples were used and distributed into eight groups according to the filling protocol.
Subsequently, the samples were sectioned into 1-mm-thick slices, and qualitatively evaluated using laser confocal scanning microscopy with inverted fluorescence Leica TCS-SPS (Leica, Mannheim, Germany). Images of the filled areas were acquired using the epifluorescence mode with absorption and emission wavelengths for Rhodamine B of 553/568 nm and for Fluo-3 of 360/449 nm, respectively, using the Leica Application Suite-Advanced Fluorescence (Leica Systems).
Samples were analyzed 10 m below the sampling surface using an objective lens with 20x magnification in a 5x5mm field of view, with a resolution of 512x512 pixels. The slices were qualitatively analyzed for each group, subgroup and thirds in which sealer penetration and the density of tags formed were observed.
Subsequently, the samples were sectioned into 1-mm-thick slices, and qualitatively evaluated using laser confocal scanning microscopy with inverted fluorescence Leica TCS-SPS (Leica, Mannheim, Germany). Images of the filled areas were acquired using the epifluorescence mode with absorption and emission wavelengths for Rhodamine B of 553/568 nm and for Fluo-3 of 360/449 nm, respectively, using the Leica Application Suite-Advanced Fluorescence (Leica Systems).
Samples were analyzed 10 m below the sampling surface using an objective lens with 20x magnification in a 5x5mm field of view, with a resolution of 512x512 pixels. The slices were qualitatively analyzed for each group, subgroup and thirds in which sealer penetration and the density of tags formed were observed.
5
Mineralization Analysis via VON KOSSA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
VON KOSSA Staining Kit (Genmed, Shanghai, China). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), Fluo-3 (Sigma, USA) dye, rhodamine-phalloidin (Sigma, MO, USA), wortmannin (WM), and β-glycerophosphate were purchased from Sigma (St. Louis, USA) respectively.
6
Scleral Fibroblast Ca2+ Dynamics Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used fluo-3 (sigma-aldrich 46393-1MG-F) to analyse the scleral fibroblast cell Ca2+ activities. Aspirate the culture media and rinse with PBS twice. Cells were given fluo-3 treatment and incubated for 30-60 min at room temperature, carried out in dark conditions. The recovery process was carried out by adding EBSS and incubated at 37°C for 30 min. Incubate for 2 min before observation. Cover with a glass cover and avoid exposure to light. Confocal laser scanning microscopy observation using 364 nm excitation with 515-560 nm emission using a confocal microscope.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!