Tissue culture medium (DMEM 01–055-1A, Biological Industries), heat-inactivated fetal bovine serum (04–121-1A, Biological Industries), antibiotic solutions (03–033-1B, Biological Industries), L-glutamine solutions (03–020-1B, Biological Industries), human recombinant insulin (01–818-1H, Biological Industries), phosphate-buffered saline (02–023-1A, Biological Industries) and Earle’s balanced salts solution (EBSS)(02–010-1A, Biological Industries). Indomethacin (I7378), dexamethasone (D4902), 3-isobutylmethylxanthine (IBMX; I7018), rosiglitazone (R2408) were obtained from Sigma-Aldrich. CYTO-ID and chloroquine (ENZ-51031, Enzo Life Science). BODIPY-C12 and Hoechst 33342 (D3835, H1399, Thermo Fisher Scientific,Inc). 3MA (M9281, Sigma-Aldrich). SMLA (SNAL-1, Protein Laboratories Rehovot Ltd).
Bodipy c12
Manufactured by Thermo Fisher Scientific
Sourced in United States
BODIPY-C12 is a fluorescent lipid probe that is useful for monitoring lipid metabolism and trafficking in cells. It consists of a BODIPY fluorescent dye covalently linked to a 12-carbon fatty acid chain. BODIPY-C12 can be used to visualize lipid droplets and other lipid-rich structures within cells.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (2 mentions)
Bodipy 558 568 c12, by Thermo Fisher Scientific (2 mentions)
Lsm 880 confocal microscope, by Zeiss (2 mentions)
Chloroquine, by Enzo Life Sciences (1 mentions)
Dmem 01 055 1a, by Sartorius (1 mentions)
Phosphate buffered saline (pbs), by Sartorius (1 mentions)
Cyto id, by Enzo Life Sciences (1 mentions)
Trypan blue reagent, by Thermo Fisher Scientific (1 mentions)
R37606, by Thermo Fisher Scientific (1 mentions)
Inverted microscope, by Olympus (1 mentions)
Lecithin, by Carl Roth (1 mentions)
Bodipy, by Thermo Fisher Scientific (1 mentions)
Hoechst, by Merck Group (1 mentions)
Nile red, by Merck Group (1 mentions)
Lsm 780 confocal microscope, by Zeiss (1 mentions)
Bodipy 493, by Thermo Fisher Scientific (1 mentions)
Beckman cytoflex s flow cytometer, by Beckman Coulter (1 mentions)
Mitosox, by Thermo Fisher Scientific (1 mentions)
Fsl confocal microscope, by Olympus (1 mentions)
18 protocols using bodipy c12
1
Adipogenic Differentiation Assay Reagents
2
Lipid Uptake Quantification in HC11 Cells
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The HC11 cell line was digested and seeded into a 96-well plate (approximately 5 × 103 cells per well). Following stable adherence and culture to 75% confluence, the cells were transfected with the plasmid and cultured for a further 24 h. Before the assay, the cells were starved in serum-free medium for 12 h, and then incubated with Bodipy-C12 (Thermo Fisher Scientific, Bedford, MA, USA) for 5 min at 37 °C without light. After quenching the extracellular fluorescence with trypan blue reagent (Gibco), the absorbance was measured by multifunctional microplate reader and recorded using an inverted fluorescence microscope (FSM-Precision, Suzhou, China), and the value was normalized to CCK8 analysis.
3
VEGF Signaling Regulation of Lipid Uptake
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Rptorfl/fl MPMECs were transduced with Ad-CMV-Null or Ad-CMV-iCre for 24 hrs. Cells were stimulated with recombinant mouse VEGF-A (300 ng/mL, R&D Systems #493-MV-005), recombinant mouse VEGF-B186 (300 ng/mL), mouse VEGFR1/Flt-1 antibody (1 μg/mL, R&D Systems, #AF471), or a combination of anti-VEGFR1 and VEGF-B for 30 hrs. For the combination, cells were pre-incubated with anti-VEGFR1 for 2 hr prior to addition of VEGF-B. Cells were then incubated with 10 μM of BODIPY FL C16 or BODIPY 500/510 C1,C12 (BODIPY-C12, Thermo #D3823) for 3 min. For imaging, cells were fixed in 4% PFA for 10 min and stained with DAPI (Invitrogen #R37606) as directed. Cells were imaged using an Olympus inverted microscope. For flow cytometry, cells were trypsinized and fixed in 4% PFA for 10 min. Flow cytometry was immediately performed as described above.
4
Quantifying Intracellular Lipid Accumulation
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BODIPY-493/503 (Thermo Fisher Scientific) and BODIPY-C12 (Thermo Fisher Scientific) were utilized to determine lipid accumulation and fatty acid uptake in cells. Cells were cultured to the desired status and washed twice with PBS. Then, BODIPY-493/503 (5 μM) or BODIPY-C12 (5 μM) was added, and the cells were incubated in the dark at 37°C. After 15 min, the dye was removed and the cells washed three times with PBS before being examined. Cells were observed using a confocal microscope. More than 4,000 cells were counted. The images were processed using ImageJ, and the fluorescent signal was collected and quantified for statistical analysis.
5
Lipid Staining Protocol Compendium
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Hoechst (Sigma), fatty acid-free BSA (PAN), Phenylmethylsulfonyl fluoride (PMSF, Sigma), BODIPY-C12 (BODIPY™ 558/568 C12, D3835, ThermoFisherScientific), BODIPY-FL-C12 (green, D3822, ThermoFisherScientific), Nile Red (Sigma), BODIPY (D3922, ThermoFisherScientific), BODIPY-Cholesterol (7356, Setareh Biotech), BODIPY-methyl-ester (CellTrace, C34556, ThermoFisherScientific), Capric acid (8797.1, Carl Roth), Glycerin-tripalmitoleate (T2630-100MG, Sigma), Lecithin (9812.1, Carl Roth).
6
Visualizing Lipid Droplets in Live Cells
7
Cellular Lipid and ROS Profiling
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Flow cytometry was utilized to determine total lipid content, cellular Reactive Oxygen Species (ROS) level, and fatty acid uptake 45 (link). Staining reagents Bodipy 493, Bodipy C12, and MitoSoX (Cat# D3922, D3822 and M36009, respectively, all purchased from Thermo Fisher) were diluted 1:1000. Following dilution, cells were stained in serum-free culture medium with the staining reagent for 30 min. After the incubation period, cells were washed twice with Phosphate Buffered Saline (PBS) to remove excess reagent. The samples were then analyzed using the Beckman CytoFLEX S flow cytometer (Beckman Coulter, Indianapolis, IN, United States) for quantitative cellular profiling.
8
Lipid Uptake Imaging in BTSC475 Cells
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BTSC475 cells were seeded in 4-well chamber slides and infected as described above. The cells were then incubated with 50nM Bodipy-C12 (Thermo Fisher Scientific, #D3822) in PBS for 15 minutes at room temperature. Subsequently, the samples were fixed with 4% paraformaldehyde in PBS, counterstained with DAPI and imaged using a FSL confocal microscope (Olympus).
9
Enterocyte Fatty Acid Uptake Assay
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Enterocytes (of fasted animals or of animals 30 min after a HFD TM) were isolated as described above. Bodipy-C12 (Molecular Probes #D3823) was pre-incubated with fatty acid–free bovine serum albumin (BSA) (2:1 M ratio) in PBS as described earlier [32] . Before resuspension with matrigel and plating, the cell pellet was incubated with Bodipy-FA conjugated with BSA (Bodipy-FA/BSA) for 5 min at 37 °C. Cells were washed twice and plated (200,000 living cells/well) in 0.5% BSA in PBS and 0.4% trypan blue (MP Biomedicals #1691049). Intracellular fluorescence was measured (bottom-read) with a microplate reader (excitation 503 nm, emission 548 nm, Spark 10M).
10
Lipid Visualization in Early Embryos
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Injection volume for all samples was maintained between 3 and 5 nl. Na-oleate (Tocris Biosciences) was used at a concentration of 1 mM. Dodecanoic acetyl CoA ammonium salt (Avanti Polar Lipids) was also used at a concentration of 1 mM. Ten micrometre solution of BODIPY-C12 (BODIPY 558/568 C12; 4,4-difluoro-5-(2-thienyl)-4-bora-3a,4a-diaza-s-indacene-3-dodecanoic acid; Molecular Probes) was prepared and injected into either the yolk or the blastodisc as mentioned in figure 3 c.
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