9 10 3h oleic acid

Manufactured by PerkinElmer

Sourced in Germany, United States

[9,10-3H]oleic acid is a radioactively labeled fatty acid compound used for research and analytical purposes. It serves as a tracer to study the metabolism, transport, and incorporation of oleic acid in various biological systems.

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Lab products found in correlation

D 2 3h glucose, by PerkinElmer (1 mentions) 5 3h d glucose, by PerkinElmer (1 mentions) D glucose, by PerkinElmer (1 mentions) 14c glucose, by PerkinElmer (1 mentions) Win55 212 2, by Bio-Techne (1 mentions) Lipidex 1000 resin, by PerkinElmer (1 mentions) Oleic acid, by PerkinElmer (1 mentions) Tri carb 2800tr, by PerkinElmer (1 mentions) Ultima gold, by PerkinElmer (1 mentions)

7 protocols using 9 10 3h oleic acid

Murine Metabolic Assessment Protocol

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2% Isoflurane, 8-0 polyprolene suture, buprenorphine 0.01 mg/kg, 2% Evan’s Blue, 1% 2, 3, 5-triphenyltetrazole (TTC, Sigma, T8877-100g), Leica microscope (Germany), biochemical analyzer, d-glucose 7 mM, insulin 10 mU/L, Oleate 0.4 mM, 1% bovine serum albumin (BSA), [9, 10]-³H-oleic acid (50 mci/L), ¹⁴C-glucose (20 mci/L), d-[2–3H] glucose (1 mci/L) and d-[5–3H] glucose (1 mci/L) were purchased from Perkin Elmer Company in the United States.

Guo Z., Wang M., Ying X., Yuan J., Wang C., Zhang W., Tian S, & Yan X. (2023). Caloric restriction increases the resistance of aged heart to myocardial ischemia/reperfusion injury via modulating AMPK–SIRT1–PGC1a energy metabolism pathway. Scientific Reports, 13, 2045.

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Cannabinoid Receptor Binding Assay

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Docosahexaenoic acid (DHA), arachidonic acid (AA), oleic acid (OA), and all other common chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA). Three of the four representative cannabinoid ligands, Δ⁹-tetrahydrocannabinol (Δ⁹-THC), CP55940 (2-[(1R,2R,5R)-5-hydroxy-2-(3-hydroxypropyl)cyclohexyl]-5-(2-methyloctan-2-yl)phenol), and SR141716A (rimonabant; 5-(4-Chlor ophenyl)-1-(2,4-dichloro-phenyl)-4-methyl-N-(piperidin-1-yl)-1H-pyrazole-3-carboxamide), were obtained from the National Institute on Drug Abuse (NIDA). The cannabinoid ligand WIN55212-2 ((R)-(+)-[2,3-Dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-napthalenylmethanone) was purchased from Tocris Bioscience (R&D Systems, Minneapolis, MN, USA). The selective adipocyte FABP (FABP4) inhibitor, BMS309403 (2-[2′-(5-Ethyl-3,4-diphenyl-1H-pyrazol-1-yl)biphenyl-3-yloxy]-acetic acid), was purchased from EMD Chemicals (San Diego, CA, USA). The [9, 10-³H] oleic acid and Lipidex 1000 resin used for the radioactive competition binding assay were from Perkin Elmer (Waltham, MA, USA).

Tyukhtenko S., Chan K., Jiang R., Zhou H., Mercier R.W., Yang D.P., Makriyannis A, & Guo J.J. (2014). Hydrogen-Bonded His93 As a Sensitive Probe for Identifying Inhibitors of the Endocannabinoid Transport Protein FABP7. Chemical biology & drug design, 85(5), 534-540.

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Lipid Uptake in Adipose Tissue

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Two experiments (either intraduodenal or intravenous infusion of TG) were performed to determine lipid uptake by adipose tissue. For intraduodenal TG infusions, CCK-KO and WT mice (n= 5 per group) maintained on LFD received a duodenal infusion of a lipid emulsion containing 4 µmoles/h butter fat and labeled with [9,10-³H]oleic acid, 1µCi/0.3 ml (Perkin Elmer, Boston, MA) 24 h after recovery from the duodenal cannulation. After the 2-h infusion, inguinal and epididymal adipose tissue was collected on dry-ice. For intravenous infusions, we used Liposyn III (intravenous fat emulsion), which it acts like a chylomicron and can transport TG directly to all tissues including adipose tissues, liver and heart [46 (link)]. The 5-h fasted mice (n=6–7 per group) received 100 µl of Lyposyn III (20% fat) mixed with (0.25 µCi, ³H-TG per mouse) over 30 min and tail blood (20 µl) was collected at 2, 4, 6, 10, 20 and 30 min post-injection. Based on previous reports [47 (link), 48 (link)], chylomicrons in the plasma are cleared in the rodents within 30 min. After 30 min, adipose tissue was collected on dry-ice. According to our published protocols [45 (link), 48 (link)], tissue lipids were extracted using the Folch method [42 (link)] and radioactivity in the plasma and tissues was measured by liquid scintillation counting.

King A., Yang Q., Huesman S., Rider T, & Lo C.C. (2015). Lipid transport in cholecystokinin knockout mice. Physiology & behavior, 151, 198-206.

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Oleic Acid Uptake in Macrophages

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MPHs were incubated with 0.4μci/ml [9,10-³H] oleic acid (PerkinElmer Life Sciences) and 100 μM unlabeled oleic acid (conjugated with BSA) in krebs-Ringer buffer(119 mM NaCl, 5 mM KCl, 2 mM CaCl₂, 2.6 mM MgSO₄,24.6 mM NaHCO₃, 2.6 mM KH₂PO₄, 10 mM HEPES, pH 7.4) for 1 h at 37°C. The supernatant was collected and incubated with 1.3 M perchloric acid. All solutions were centrifuged at 16,000 g for 10 minutes, and the supernatant was neutralized with 2 M KOH and 0.6 M MOPS. 3ml scintillation was added into solution and measure [³H] radioactivity.

Yang Z., Yu Y., Sun N., Zhou L., Zhang D., Chen H., Miao W., Gao W., Zhang C., Liu C., Yang X., Wu X, & Gao Y. (2020). Ginsenosides Rc, as a novel SIRT6 activator, protects mice against high fat diet induced NAFLD. Journal of Ginseng Research, 47(3), 376-384.

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Oleic Acid Oxidation Assay in Hepatocytes

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The assays were performed as previously described at Soochow Pukang Biotechnology Co., LTD.^{35 (link),47 (link)} Briefly, primary hepatocytes infected with the indicated adenovirus were incubated with 0.4 μCi/ml [9,10-³H]oleic acid (Perkin Elmer Life Sciences) and 100 μm unlabeled oleic acid (conjugated with BSA) in Krebs-Ringer buffer (119 mM NaCl, 5 mM KCl, 2 mM CaCl₂, 2.6 mM MgSO₄, 24.6 mM NaHCO₃, 2.6 mM KH₂PO₄, 10 mM HEPES, pH 7.4) for 3 h at 37 °C. Chloroform was used to extract the unoxidized oleic acid. Supernatants were collected and incubated with 1.3 M perchloric acid. All the solutions were then centrifuged at 16,000 × g for 10 min. Supernatants were neutralized with 2 M KOH, 0.6 M MOPS. Next, 3 mL of scintillation liquid was then added and [³H] radioactivity was measured.

Zhang L., Chen J., Yang X., Shen C., Huang J., Zhang D., Liu N., Liu C., Zhong Y., Chen Y., Tang K., Guo J., Cui T., Duan S., Li J., Huang S., Pan H., Zhang H., Tang X., Chang Y, & Gao Y. (2024). Hepatic Zbtb18 (Zinc Finger and BTB Domain Containing 18) alleviates hepatic steatohepatitis via FXR (Farnesoid X Receptor). Signal Transduction and Targeted Therapy, 9, 20.

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Oleic Acid Uptake in Adenovirus-Infected Cells

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Adenovirus-infected MPHs were incubated with 0.4 µCi/mL [9,10-³ H] oleic acid (PerkinElmer Life Science) and 100 µM unlabelled oleic acid (conjugated with bovine serum albumin (BSA)) in Krebs-Ringer buffer (119 mM NaCl, 5 mM KCl, 2 mM CaCl₂, 2.6 mM MgSO₄, 24.6 mM NaHCO₃, 2.6 mM KH₂PO₄, 10 mM HEPES, pH 7.4) for 1 hour at 37°C. Supernatants were collected and incubated with 1.3 M perchloric acid. All samples were next centrifuged at 16 000× g for 10 min. Supernatants were neutralised with 2 M KOH, 0.6M MOPS. Scintillation liquid (3 mL) was then added, and [³H] radioactivity was measured.

Sun N., Shen C., Zhang L., Wu X., Yu Y., Yang X., Yang C., Zhong C., Gao Z., Miao W., Yang Z., Gao W., Hu L., Williams K., Liu C., Chang Y, & Gao Y. (2020). Hepatic Krüppel-like factor 16 (KLF16) targets PPARα to improve steatohepatitis and insulin resistance. Gut, 70(11), 2183-2195.

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Oleate Uptake Kinetics in Hepatocytes

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Uptake of [ 3 H]oleate was measured in confluent HepG2 cells and rat primary hepatocytes as previously described [48, 49] . Briefly, 0.68µCi of [9,10-3 H]oleic acid (54.6 Ci/mmol; Perkin Elmer) was mixed with 50µM of non-radioactive oleate (Sigma-Aldrich) and dissolved in a 173µM BSA solution free of fatty acids. Cells were incubated with this oleate/BSA solution for 10min at 37 °C. Uptake was stopped by removal of the oleate/BSA solution followed by the addition of ice-cold 1x PBS (5ml) containing 200µM of phloretin and 0.1% BSA (wt/v). After a 2min incubation, cells were washed six times with ice-cold 1x PBS. Cells were then lysed with 2M NaOH and aliquots of the lysate were used for protein concentration and radioactivity measurements. Radioactivity was measured in a TRI Carb 2800TR liquid scintillation counter after the addition of 10ml Ultima-Gold (Perkin Elmer). Data were presented as number of counts per minute (CPM) of [ 3 H] per µg of protein.

Lounis M.A., Lalonde S., Rial S.A., Bergeron K.F., Ralston J.C., Mutch D.M, & Mounier C. (2017). Hepatic BSCL2 (Seipin) Deficiency Disrupts Lipid Droplet Homeostasis and Increases Lipid Metabolism via SCD1 Activity. Lipids, 52(2).

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