Cells were washed twice with PBS and fixed with paraformaldehyde (PFA, 4% in PBS) for 30 min. Cells were then washed twice with PBS and once with distilled water, and stained with Oil Red O dye (6:4, 0.6% Oil Red O dye in water; Sigma (Sintra, Portugal)) for 1 h at room temperature, and then washed three times with water. Finally, Oil Red O was dissolved in 200 μL iso- propanol, and absorbance was measured at 500 nm using the Multiskan EX (Thermo Scientific; Waltham, MA, USA).
Oil red o dye
Manufactured by Merck Group
Sourced in United States, Germany, Portugal
Oil Red O dye is a fat-soluble azo dye used in histology and cytology for the detection of neutral triglycerides and lipids. It stains lipids and lipoproteins a red color, allowing for their visualization and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Insulin, by Merck Group (10 mentions)
3 isobutyl 1 methylxanthine, by Merck Group (6 mentions)
Dexamethasone (dex), by Merck Group (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Oil red o, by Merck Group (3 mentions)
Alcian blue, by Merck Group (3 mentions)
Rosiglitazone, by Merck Group (2 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (2 mentions)
Antibodies against pparγ, by Santa Cruz Biotechnology (2 mentions)
Eclipse ti2 u, by Nikon (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Paraformaldehyde (pfa), by Merck Group (2 mentions)
Stempro chondrogenic, by Thermo Fisher Scientific (2 mentions)
Ckx53, by Olympus (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Trypsin, by Merck Group (2 mentions)
Indomethacin, by Merck Group (2 mentions)
Multiskan ex, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Lonza (1 mentions)
Protein marker, by Bio-Rad (1 mentions)
65 protocols using oil red o dye
1
Oil Red O Staining for Lipid Quantification
2
Adipogenesis Pathway Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The culture medium Dulbecco's modified Eagle's medium (DMEM) used in all the experiments was purchased from Lonza. The triacylglycerol assay kit was purchased from Abnova. NBD-ceramide (NBD-N-hexanoyl-D-erythro-sphingosine) was from Cayman Chemicals. Dexamethasone, rosiglitazone, insulin, 3-isobutyl-1-methylxanthine (IBMX), and the Oil Red O dye were obtained from Sigma-Aldrich. Nitrocellulose membranes, protein markers, and BCA assay reagents were purchased from Bio-Rad. Fetal bovine serum (FBS) and newborn calf serum (NCS) were from GIBCO. The ELISA kit for determination of leptin was purchased from Peprotech. The PPARγ antibody was supplied by Cell Signaling. The GAPDH antibody, nontargeting (negative) siRNA, and ceramide kinase (CerK) siRNA were purchased from Santa Cruz Biotechnology. The CerK antibody was from Calbiochem or Abgent. The rest of chemicals and reagents used in this work were of the highest grade available.
3
Lipid Droplet Staining in ESCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Naïve and primed ESCs were fixed with 4% PFA at room temperature for 10min, washed twice with PBS and stained with Oil Red O dye (Sigma) for 10min at rt. Alternatively, lipid droplets were stained using BODIPY 493/503 (Molecular Probes) for 15min on a rocking platform at rt. Pictures were taken using a fluorescent microscope (Leica). Lipid droplet analysis at in vivo post-implantation stage has proven to be difficult.
4
Histological Assessment of Liver Pathology
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Portions of fresh liver were formalin-fixed and paraffin-embedded and subsequently subjected to hematoxylin and eosin (H&E) and Sirius red staining. IHC was conducted as previously described. Briefly, paraffin-embedded liver sections were deparaffinized and rehydrated, followed by incubation with anti-F4/80 antibody (Cell Signaling Technology, Danvers, MA, USA). For Oil red O staining, frozen liver portions were stained with Oil red O dye (Sigma, St. Louis, MO, USA) according to the manufacturer’s protocols. Three independent visual fields were randomly selected from each slide at 20× magnification and analyzed using ImageJ software. Pathological features were estimated according to the NAFLD activity score (NAS), Sirius red area, Oil red O staining area, and F4/80 staining intensity.
5
Characterization of Gold Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We purchased MC (Mw =14,000 g mol−1), phosphate-buffered saline (PBS, pH 7.4), Oil Red O dye, dexamethasone, insulin, isobutylmethylisobutylxanthine (IBMX), leptin from mouse (>98%), and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) from Sigma-Aldrich Co. (St Louis, MO). CellTiter 96® AQueous One Solution Cell Proliferation Assay kit for the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was purchased from Promega (Madison, WI). Bare gold NPs (A11-20, size: 20 nm) were purchased from Nanopartz™ Inc (Loveland, Colorado). For transmission electron microscopy (TEM, JEOL JEM-1400) analysis, a drop of gold NP solution was allowed to air-dry onto a Formvar-carbon-coated 200-mesh copper grid. TEM images were imaged on a JEOL-1400 transmission electron microscope operating at an accelerating voltage of 100 kV. The UV–vis absorption spectra of the gold NP solutions that were prepared under deionized water were measured using a UV–vis spectrophotometer (Tecan’s Sunrise; Tecan Trading AG, Männedorf, Switzerland).
6
Oil Red O Lipid Staining Protocol
7
Adipokine Profiling of Mouse Adipocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteome Profiler Mouse Adipokine Array Kit (Catalog # ARY013) was purchased from R&D Systems (Minneapolis, MN, USA). Dulbecco’s modified Eagle’s medium (DMEM), and fetal bovine serum (FBS) were purchased from Thermo Fisher Scientific (Carlsbad, CA, USA). Insulin, 3-isobutyl-1-methylxanthine (IBMX), dexamethasone (DEX), Oil Red O dye, and chloroquine (CQ) were purchased from Sigma–Aldrich (St. Louis, MO, USA).
8
Adipocyte Differentiation Assay Protocol
9
Quantifying Intracellular Triglycerides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intracellular triglycerides, a major component of milk fat, were determined using oil red O dye (Sigma-Aldrich). The MAC-T cells were fixed using 10% formalin at 25 °C for 1 h and washed with 60% isopropanol. After completely drying, the oil red O working solution was used to stain the cells for 10 min. The unbound dye was removed using deionized distilled water, whereafter the stained cells were captured and imaged using a Nikon Eclipse Ti2-U and a Nikon Eclipse Ts2R camera. The oil red O staining areas were quantified using the Image J software, version 1.53.
10
Quantification of Lipid Accumulation in 3T3-L1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lipid accumulation was detected in 3T3-L1 cells after staining with Oil red-O dye, as described in previous reports [7 (link)]. Briefly, cells were fixed with 4% formaldehyde for 60 min and washed three times with distilled water, after which they were incubated with 0.5% Oil red-O dye (Sigma-Aldrich Co.) in 100% isopropanol for 30 min at room temperature. After washing three times with distilled water, the stained fat droplets in the adipocytes were observed microscopically at 100× magnification (Leica Microsystems, Wetzlar, Germany). Spectrophotometric analysis of the stain was performed by dissolving the stained lipid droplets in isopropanol. Finally, the absorbance was measured at 510 nm using a Molecular Devices VERSA max Plate reader.
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!