Cells or tissues were washed 3 times with PBS and then fixed with formaldehyde. Fixed cells or tissues were stained with oil red O and hematoxylin and eosin. For fluorescent lipid droplet staining, AdipoRedTM assay reagent (Lonza) was used, and the nuclei were stained with a DAPI DNA stain.
Adipored assay reagent
Manufactured by Lonza
Sourced in United States, Switzerland
The AdipoRed Assay Reagent is a fluorescent dye designed to quantify adipocyte differentiation in cell culture models. It allows for the measurement of lipid accumulation, a key indicator of adipogenesis.
Automatically generated - may contain errors
Lab products found in correlation
3 isobutyl 1 methylxanthine, by Merck Group (6 mentions)
Dexamethasone (dex), by Merck Group (6 mentions)
Insulin, by Merck Group (4 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (4 mentions)
Oil red o staining kit, by Abcam (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Hoechst 33342, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
2 n 7 nitrobenz 2 oxa 1 3 diazol 4 yl amino 2 deoxy d glucose 2 nbdg, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
α glucosidase, by Merck Group (2 mentions)
Acarbose, by Merck Group (2 mentions)
Insulin from bovine pancreas, by Merck Group (2 mentions)
Fetal bovine calf serum, by GE Healthcare (2 mentions)
Penicillin streptomycin, by GE Healthcare (2 mentions)
Glomax multi microplate multimode reader, by Promega (2 mentions)
96 well plate, by Dutscher (2 mentions)
Spectramax i3, by Molecular Devices (2 mentions)
Orca flash4.0 hamamatsu digital camera c11440, by Hamamatsu Photonics (2 mentions)
57 protocols using adipored assay reagent
1
Lipid Droplet Staining in Cells
2
Triglyceride Quantification in HepaRG Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Triglyceride accumulation in HepaRG cells after oleic acid treatment was quantified using AdipoRedTM Assay Reagent (Lonza, Walkersville, MD, United States). Briefly, the cells were washed in PBS and AdipoRedTM Assay Reagent was added. After a 10-min incubation at room temperature, the fluorescence intensity was quantified using a SynergyTM H4 hybrid multi-mode microplate reader (BioTek, Winooski, VT, United States).
3
Cell Culture Media and Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dulbecco’s modified Eagle’s medium (DMEM) was purchased from Gibco (Grand Island, NY, USA). Bovine calf serum (BCS), fetal bovine serum (FBS), penicillin (100 units/mL)/streptomycin (100 μg/mL), and Trizol were purchased from Invitrogen (Carlsbad, CA, USA). Acarbose, dexamethasone, fluorescein, insulin, oleic acid, Trolox, 3-isobutyl-1-methylxanthine, α-glucosidase, and β-phycoerythrin were obtained from Sigma Aldrich (St. Louis, MO, USA). AdipoRedTM assay reagent was purchased from Lonza (Basel, Switzerland). 2-[N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG) was obtained from Thermo Fisher Scientific (Carlsbad, CA, USA). Chemicals, unless otherwise stated, were purchased from Sigma Aldrich.
4
Adipocyte Differentiation and Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Diethyl ether, Oil red O, simvastatin (Simv), insulin, dexamethasone, 3-isobutyl-1-methylxanthine (IBMX), TNF-α and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma (St. Louis, MO). Omega-3 was purchased from GC Rieber (AS, Kristiansand, Norway). Antibodies against vascular cell adhesion molecule-1 (VCAM-1) and P-selectin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). AdipoRedTM assay reagent was purchased from Lonza (Walkersville, MD).
5
Quantifying MSC Differentiation Potential
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Unless indicated otherwise, MSCs (1000 cells/cm2) at passage 2 were maintained in 5% oxygen and their capacity for adipogenic and osteogenic differentiation was quantified as described previously [16 (link)] with the following exceptions. Adipogenic induction media (AIM) consisted of α-MEM supplemented with 10% rabbit serum, 10−8 M dexamethasone, 20 mM ETYA, and 25 mg/ml insulin. Lipid accumulation was quantified by staining the monolayer with AdipoRedTM Assay Reagent (Lonza) for 10 min and measuring fluorescence using a SpectraMax fluorescent plate reader (excitation 485 nm; emission 572 nm). Adipo-induction was normalized to cell number, which was determined by staining the monolayer with 4', 6-diamidino-2-phenylindole (DAPI) or direct cell counting. Osteogenic induction media (OIM) consisted of high-glucose DMEM supplemented with 10% FBS, 50 µg/ml L-ascorbic-2-phosphate, 10 mM β-glycerolphosphate, and 10−8 M dexamethasone. Mineralization was quantified by extracting calcium from monolayers using 10% cetylpyridinium chloride and measuring absorbance at 562 nm. Osteo-induction was also normalized to cell number.
6
Quantification of Lipid Droplet Accumulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded on a chamber slide, and adipogenesis differentiation was induced in the presence or absence of Sq and HH-Sq as described above. Lipid droplets (LDs) formed after differentiation were stained by fluorescent neutral lipid dye 4,4-difluoro-1,3,5,7,8-pentamethyl-4-bora-3a,4a-diaza-s-indacene (BODIPY 493/503; Invitrogen, Eugene, OR, United States), mounted with ProLongTM Diamond Antifade Mountant with DAPI (Invitrogen, Eugene, OR, United States) for staining nucleus, and fluorescence images were obtained by using Leica TCS SP8 confocal microscope. For quantification of LD accumulation, cells were seeded in a 96-well plate and induced adipogenesis differentiation. After washing with PBS, 5 μL of AdipoRedTM Assay Reagent (Lonza) was added in each well containing 200 μL PBS and incubated for 10 min at room temperature. Fluorescence with excitation at 485 nm and emission at 572 nm was measured by using a microplate reader (Varioskan LUX).
7
Adipogenesis Assay in HepG2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HepG2 cells were seeded into 6-well (30,000/cm2) plates for lipid accumulation assays. The cells were grown to confluence using Dulbec47co’s Modified Eagle Medium (DMEM) low glucose media (Gibco/Thermo Fisher Scientific, Waltham, MA, USA), 1% Penicillin-Streptomycin-Neomycin Antibiotic Mixture (PSN, Thermo Fisher Scientific), and fetal bovine serum (FBS, Thermo Fisher Scientific). Then the cells were switched to an FBS-free media (DMEM and PSN only) for 48 h, followed by treatment of FBS-free media with 1% human plasma sample (PCOS Day 0 (PCOS-D0), PCOS Day 7 (PCOS-D7), CON Day 0 (CON-D0) or CON Day 7 (CON-D7)) added for an additional 48 h. Plasma samples added were from baseline draws on the respective days with a minimum of 18 h post previous ingestion of WPI supplementation. Plasma from 6 different participants from each group of the clinical part of this study were used for these experiments (one plasma source per well). Once HepG2 cells had been treated with human plasma for 48 h, lipid accumulation was measured using AdipoRed™ Assay Reagent (Lonza, Walkersville, MD, USA) per manufacturer’s instructions for a 6-well plate [47 ].
8
Adipogenic Differentiation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen cells (passage 3–4) were revived from frozen in fresh growth medium and plated in triplicate 12‐well plates (5000 c/cm2). At confluence, the medium was switched to either low glucose maintenance medium (controls; DMEM‐low glucose, 10% FBS, 2 mM L‐glutamine, 5 μg/ml gentamycin) or commercial differentiation‐inducing medium (differentiated; Stem‐Pro Adipogenesis Differentiation kit, cat. A10070‐01; Life Technologies; Carlesbad, CA). After 12–14 days, control and differentiated cells were stained for lipid with Oil Red O for 15 min, followed by multiple rinses with ddH2O as previously described 24 . Oil Red O was then extracted with isopropanol and absorbance measured at 569 nm. Differentiation was also quantified using AdipoRed Assay Reagent (Lonza, Walkersville, MD) per manufacturer's instructions and fluorescence was detected at (485EX/572EM). Differentiation values are reported as a percent of undifferentiated, control cells.
9
Epinephrine-Induced Lipid Accumulation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 1.0 × 104 HaCaT were seeded in 96-well plates, grown for 48 h, and then treated with the FcHEx or ICI-118,551 as a positive control for an additional 24 h. The cells were incubated with 10 µM of epinephrine for 24 h, and the lipid accumulation was detected via the addition of the AdipoRed assay Reagent (Lonza, Basel, Switzerland). The fluorescence at 485 nm was read using the Victor3 plate reader system, and the content of the epidermal lipid was normalized relative to the cell density determined by crystal violet staining.
10
Adipogenic Differentiation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
On day 5 of differentiation, cells were replated μClear plates as described above. On day 7, cells were washed three times and treated with given inhibitor combinations for 2 h. Cells were then washed two times and incubated with 100 μl (40x dilution in PBS) adipoRed assay reagent (Lonza PT 7009) for 15 min and read on a microplate reader using Ex = 485 nm, Em = 535 nm.
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!