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Adipored assay reagent kit

Manufactured by Lonza
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The AdipoRed Assay Reagent kit is a fluorescence-based reagent used for the quantitative determination of adipogenesis in cell-based assays. The kit provides a direct and reliable method for measuring lipid accumulation in cells.

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Lab products found in correlation

Victor3, by PerkinElmer (9 mentions) Free glycerol reagent, by Merck Group (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Cholesterol assay kit, by Abcam (1 mentions) Victor3 plate reader, by PerkinElmer (1 mentions) 3 isobutyl 1 methylxanthine ibmx, by Merck Group (1 mentions) Oil red o working solution, by Merck Group (1 mentions) Enhanced chemiluminescence ecl reagent, by Advansta (1 mentions) Insulin, by Merck Group (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions)

11 protocols using adipored assay reagent kit

1

Quantification of Lipid Metabolism in Adipocytes

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The collected blood was centrifuged at 3,000 x g at 4˚C for 15 min to obtain serum. In serum and/or mature 3T3-L1 cells, TG and TC contents were measured with an AdipoRed assay reagent kit (Lonza Group, Ltd.) or Cholesterol assay kit (Abcam), respectively, in accordance with the manufacturers' instructions. Fluorescence was measured following 10 min of incubation on a Victor3 plate reader (PerkinElmer, Inc.) with the following settings: Excitation (Ex) 485/Emission (Em) 572 nm (for TG) and Ex538/Em587 (for TC).
Differentiated 3T3-L1 adipocytes were serum-starved for 2 h and treated with AZD1208 (cat. no. HY-15604; MedChemExpress) or isoproterenol, a known lipolysis inducer, for 3 h. Culture medium was obtained and used to measure glycerol contents with a free glycerol reagent (Sigma-Aldrich; Merck KGaA) according to the manufacturer's instructions. The absorbance was determined at 540 nm using a microplate reader.
Li J., Gong L, & Xu Q. (2021). Purinergic 2X7 receptor is involved in adipogenesis and lipid degradation. Experimental and Therapeutic Medicine, 23(1), 81.
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2

Cellular Lipid Quantification via AdipoRed

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On day 8 of differentiation, cellular lipid contents were measured with AdipoRed assay reagent kit in consonance with the manufacturer's instructions (Lonza, Basel, Switzerland). Fluorescence was measured after a 10 minutes incubation on Victor3 (PerkinElmer, Waltham, MA, USA) with excitation at 485 nm and emission at 572 nm.
Park Y., Obiang‐Obounou B.W., Lee K., Choi J, & Jang B. (2018). AZD1208, a pan‐Pim kinase inhibitor, inhibits adipogenesis and induces lipolysis in 3T3‐L1 adipocytes. Journal of Cellular and Molecular Medicine, 22(4), 2488-2497.
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3

Lipid Content Measurement in 3T3-L1 Cells

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On day eight of differentiation, lipid content in control or tanshinone IIA-treated 3T3-L1 cells was measured using a commercially available AdipoRed Assay Reagent kit according to the manufacturer’s instructions (Lonza, Basel, Switzerland). After a 10 min incubation, fluorescence was measured on Victor3 (Perkin Elmer, Waltham, MA, USA) with an excitation at 485 nm and an emission at 572 nm.
Park Y.K., Obiang-Obounou B.W., Lee J., Lee T.Y., Bae M.A., Hwang K.S., Lee K.B., Choi J.S, & Jang B.C. (2017). Anti-Adipogenic Effects on 3T3-L1 Cells and Zebrafish by Tanshinone IIA. International Journal of Molecular Sciences, 18(10), 2065.
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4

Quantification of Intracellular Triglycerides

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On D8 of differentiation, intracellular TG content in control or β3-AR shRNA-transfected 3T3-L1 cells were quantified using AdipoRed assay reagent kit (Lonza, Basel, Switzerland) according to the manufacturer’s protocol. Fluorescence intensity was quantified with excitation and emission at 485 and 572 nm, respectively, using Victor3 (Perkin Elmer, Boston, MA, USA).
Roshanzadeh A., Yadav A.K., Pydi S.P., Kimura T, & Jang B.C. (2022). Expression and Role of β3-Adrenergic Receptor during the Differentiation of 3T3-L1 Preadipocytes into Adipocytes. Biology, 11(5), 772.
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5

Cudrania Tricuspidata Root Extract Protocol

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Cudratricusxanthone A (CTXA) was isolated from the roots of Cudrania tricuspidata Bureau as reported previously [26 (link)]. Enhanced chemiluminescence (ECL) reagent was bought from Advansta (Menlo Park, CA, USA). 3-isobutyl-1-methylxanthine (IBMX), dexamethasone, and insulin were purchased from Sigma (St. Louis, MO, USA). Adipo-red assay reagent kit was bought from Lonza (Basel, Switzerland). Free Glycerol Reagent and Oil Red O working solution was obtained from Sigma (St. Louis, MO, USA). Pierce BCA Protein Assay Kit was bought from Thermo Scientific (Rockford, IL, USA). Antibodies used in this study are listed in Table S1.
Kwon H.S., Jeong G.S, & Jang B.C. (2021). Cudratricusxanthone A Inhibits Lipid Accumulation and Expression of Inducible Nitric Oxide Synthase in 3T3-L1 Preadipocytes. International Journal of Molecular Sciences, 22(2), 505.
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6

Intracellular Triglyceride Quantification

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Cells were similarly grown under the above-mentioned differentiation conditions. On day 8 of differentiation, intracellular TG content in control or CTXA-treated 3T3-L1 cells was measured using a commercially available Adipo-red assay reagent kit according to the manufacturer’s instructions (Lonza, Basel, Switzerland). Fluorescence was measured on Victor3 (Perkin Elmer, Waltham, MA, USA) with an excitation at 485 nm and emission at 572 nm. Data are mean ± SE of three independent experiments.
Kwon H.S., Jeong G.S, & Jang B.C. (2021). Cudratricusxanthone A Inhibits Lipid Accumulation and Expression of Inducible Nitric Oxide Synthase in 3T3-L1 Preadipocytes. International Journal of Molecular Sciences, 22(2), 505.
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7

Measuring Intracellular Triglycerides in 3T3-L1 Cells

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On D8 of differentiation, intracellular TG content in control or pazopanib-treated 3T3-L1 cells was measured with AdipoRed assay reagent kit in consonance with the manufacturer’s instructions (Lonza, Basel, Switzerland). Fluorescence intensity was quantified with excitation and emission at 485 and 572 nm, respectively, using Victor3 (Perkin Elmer, Waltham, MA, USA).
Yadav A.K, & Jang B.C. (2021). Inhibition of Lipid Accumulation and Cyclooxygenase-2 Expression in Differentiating 3T3-L1 Preadipocytes by Pazopanib, a Multikinase Inhibitor. International Journal of Molecular Sciences, 22(9), 4884.
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8

Adipose Lipid Content Measurement

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After 8 days of treatment, lipid content was measured using a commercially available AdipoRed Assay Reagent kit according to the manufacturer's instructions (Lonza). After a 10-min incubation, the plates were placed in a Victor3 (Perkin Elmer), and fluorescence was measured with an excitation wavelength of 485 nm and an emission wavelength of 572 nm.
Park Y.K., Lee J., Hong V.S., Choi J.S., Lee T.Y, & Jang B.C. (2014). Identification of KMU-3, a Novel Derivative of Gallic Acid, as an Inhibitor of Adipogenesis. PLoS ONE, 9(10), e109344.
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9

Intracellular Triglyceride Quantification

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On day 8 of differentiation, intracellular TG contents were quantified using the AdipoRed Assay Reagent kit in accordance with the manufacturer’s instructions (Lonza, Basel, Switzerland). Fluorescence was defined after a 10 min incubation on Victor3 (Perkin Elmer, Waltham, MA, USA) with excitation at 485 nm and emission at 572 nm.
Park Y.K, & Jang B.C. (2023). The Receptor Tyrosine Kinase c-Met Promotes Lipid Accumulation in 3T3-L1 Adipocytes. International Journal of Molecular Sciences, 24(9), 8086.
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10

Quantifying Intracellular Lipid Content

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Intracellular lipid content of vehicle or tanshinone I-treated 3T3-L1 cells were measured using a commercially available AdipoRed Assay Reagent kit according to the manufacturer's instructions (Lonza, Walkersville, MD, USA).
After a 10 min incubation, fluorescence was measured on a Victor3 (Perkin Elmer Inc., Waltham, MA, USA) with excitation at 485 nm and emission at 572 nm. TG content was expressed as a percentage of vehicle control.
Kwon H., & Jang B. (2020). Tanshinone I, an Active Ingredient of Salvia miltiorrhiza, Inhibits Differentiation of 3T3-L1 Preadipocytes and Lipid Accumulation in Zebrafish. Journal of Korean Medicine for Obesity Research, 20(2), 109-121.
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