Palmitate

Manufactured by Merck Group

Sourced in United States, Germany, Macao, United Kingdom, Sao Tome and Principe, France, Japan, China

Palmitate is a type of laboratory equipment used for research and analysis purposes. It is a fatty acid compound that serves as a common precursor for various biological processes. Palmitate is utilized in various scientific applications, including cell culture studies, biochemical assays, and metabolic research. The core function of Palmitate is to provide a standardized and reliable source of this essential fatty acid for experimental and analytical purposes.

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Lab products found in correlation

Oleate, by Merck Group (46 mentions) Bovine serum albumin (bsa), by Merck Group (39 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (39 mentions) Fatty acid free bsa, by Merck Group (17 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (16 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (14 mentions) Insulin, by Merck Group (11 mentions) Lipopolysaccharides (lps), by Merck Group (10 mentions) Sodium palmitate, by Merck Group (10 mentions) Linoleate, by Merck Group (9 mentions) Antimycin a, by Merck Group (8 mentions) Stearate, by Merck Group (8 mentions) Metformin, by Merck Group (7 mentions) Glucose, by Merck Group (7 mentions) Oligomycin, by Merck Group (6 mentions) Elaidate, by Merck Group (6 mentions) Vaccenate, by Merck Group (6 mentions) Mitotempo, by Merck Group (6 mentions) Bovine serum albumin (bsa), by Thermo Fisher Scientific (6 mentions) Fatty acid free bovine serum albumin, by Merck Group (6 mentions)

284 protocols using palmitate

Bovine Serum Albumin-Conjugated Palmitate Protocol

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Bovine serum albumin (BSA)-conjugated palmitate was prepared by following the procedure as previously described with some modifications (Gao et al., 2004 (link); Li et al., 2018 (link)). Briefly, a stock of 200 mM palmitate (NU-CHEK PREP, INC., Elysian, MN) in 100% EtOH was mixed with 3.3% fatty acid-free BSA solution (MilliporeSigma, Burlington, MA) to a working concentration of 1.7 mM by heating at 40°C for 1 h (the molar ratio of palmitate to BSA was 3.4). The same volume of EtOH mixed with 3.3% fatty acid-free BSA was used as control. The BSA or BSA-conjugated palmitate stock solution was then dissolved in DMEM containing 50 μM carnitine, 10 mM HEPES (pH 7.3), 26 mM NaHCO3, and 1 μg/ml gentamicin, to a final concentration of 0.5 mM palmitate (1% BSA).
After 5 days of differentiation, myotubes were treated with vehicle [VEH; phosphate buffered saline (PBS)] or 5 μM imoxin (EMD Millipore, Burlington, MA) in the absence or presence of 0.5 mM BSA-conjugated palmitate for 24 h. To evaluate insulin action, 50 nM insulin was added to the myotubes 15 min before harvest.

Eo H, & Valentine R.J. (2022). Saturated Fatty Acid-Induced Endoplasmic Reticulum Stress and Insulin Resistance Are Prevented by Imoxin in C2C12 Myotubes. Frontiers in Physiology, 13, 842819.

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Palmitate Treatment of RAW 264.7 Cells

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RAW 264.7 cells (immortalized mouse monocyte cells) were obtained from American Type Culture Collection (ATTC, Manassas, VA, USA) and were cultured in Dulbecco’s modified Eagle medium (WG, Gyeongsan, Korea) supplemented with 10% fetal bovine serum (WG, Gyeongsan, South Korea) and 1% antibiotic-antimycotic (Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C with 5% CO₂. Used cells were confirmed to be free from mycoplasma. For preparation of palmitate treatment, stock solution of 100 mM palmitate (Sigma-Aldrich, St. Louis, MO, USA) was dissolved with 0.1 M NaOH by heating at 70 °C in a hot water bath without vortex. 5% BSA solution with fatty acid free (Sigma-Aldrich, St. Louis, MO, USA) was prepared with H₂O and filtered through 0.22 μm syringe filter. By mixing two solutions, 5 mM palmitate-BSA was conjugated in a 55 °C water bath for 10–15 minutes, and cooling down before use. In some cases, 10 μM sulfosuccinimidyl oleate (SSO) (Cayman, Ann Arbor, MI, USA) or 80 μM dynasore (Sigma-Aldrich, St. Louis, MO, USA) was pre-treated for 10 min before palmitate treatment.

Kim S.Y., Jeong J.M., Kim S.J., Seo W., Kim M.H., Choi W.M., Yoo W., Lee J.H., Shim Y.R., Yi H.S., Lee Y.S., Eun H.S., Lee B.S., Chun K., Kang S.J., Kim S.C., Gao B., Kunos G., Kim H.M, & Jeong W.I. (2017). Pro-inflammatory hepatic macrophages generate ROS through NADPH oxidase 2 via endocytosis of monomeric TLR4–MD2 complex. Nature Communications, 8, 2247.

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Metabolic Effects of Palmitate and Arachidonic Acid in ob/ob Mice

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Palmitate and arachidonic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA) and Nu-chek Prep, Inc. (Elysian, MN, USA), respectively. Palmitate- and arachidonic acid-bovine serum albumin (BSA) solutions were prepared by dissolving Palmitate in ethanol and then mixing it with fatty acid-free BSA (2% wt/vol in water; Sigma-Aldrich) at 37 °C on a shaker for 2 h. We obtained 7-week-old male ob/ob mice and age-matched lean mice (C57BL/6J) from the Animal Center of SLC, Inc. (Hamamatsu, Shizuoka, Japan). The mice were housed in individual cages at 22 ± 2 °C with a 12-h light-dark cycle. After overnight fasting, the liver was removed from each mouse and used for western blot analysis. All animal experiments were approved by the institutional animal care and use committee of the Korea Center for Disease Control and Prevention (KCDC-015-11-2A). The methods were carried out in accordance with the approved guidelines.

Hwang J.Y., Lee H.J., Go M.J., Jang H.B., Choi N.H., Bae J.B., Castillo-Fernandez J.E., Bell J.T., Spector T.D., Lee H.J, & Kim B.J. (2018). Genome-wide methylation analysis identifies ELOVL5 as an epigenetic biomarker for the risk of type 2 diabetes mellitus. Scientific Reports, 8, 14862.

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Adipocyte and Macrophage Isolation and Culture

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3T3-L1 murine pre-adipocytes (ATCC), human pre-adipocytes from Simpson-Golabi-Behmel syndrome (SGBS) were propagated and differentiated according to standard procedures (Yeop Han et al., 2010 (link); Lin et al., 2005 (link)), except that cells were differentiated and propagated in DMEM containing 5 or 25 mmol/L glucose (Hyclone), and media were changed daily.
thioglycollate (TG)-elicited peritoneal macrophages and RPM were obtained from the peritoneal cavities from several mouse strains. To obtain resident macrophages, PBS was injected intraperitoneally, and cells were collected immediately. To obtain TG-elicited macrophages, thioglycollate (BD) was injected intraperitoneally and cells were collected after 4 days. To remove non-adherent peritoneal cells from resident and TG-elicited peritoneal macrophages, the plates were washed extensively after adherence of macrophages to the plates for 60 minutes.
Palmitate (16:0) (Sigma) was conjugated with albumin, as described previously (Yeop Han et al., 2010 (link)). Briefly, Palmitate was first dissolved in NaOH (100 mmol/L) and conjugated with fatty acid-free albumin (Sigma) at a molar ratio of 3:1 (Palmitate/albumin).

Han C.Y., Kang I., Harten I.A., Gebe J.A., Chan C.K., Omer M., Alonge K.M., den Hartigh L.J., Kjerulf D.G., Goodspeed L., Subramanian S., Wang S., Kim F., Birk D.E., Wight T.N, & Chait A. (2020). Adipocyte-Derived Versican and Macrophage-Derived Biglycan Control Adipose Tissue Inflammation in Obesity. Cell reports, 31(13), 107818.

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Palmitate Treatment of INS-1 Cells

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Palmitate (Sigma-Aldrich, St Louis, MO) was added to the INS-1 cells by conjugating Palmitate with fatty acid-free bovine serum albumin (BSA), and the stock solution of Palmitate was prepared as described previously
[21 (link)]. Briefly, A 5 mM Palmitate/5% BSA (5 mM PA) stock solution was prepared by mixing 1 ml 100 mM Palmitate with 19 ml 5.26% BSA in a 55°C water bath. During experiment, 5 mM PA was diluted in RPMI 1640 without fetal bovine serum (FBS) to desired concentrations (Palmitate: BSA: molar ratio of 6.6:1).

Huang S., Zhu M., Wu W., Rashid A., Liang Y., Hou L., Ning Q, & Luo X. (2014). Valproate pretreatment protects pancreatic β-cells from palmitate-induced ER stress and apoptosis by inhibiting glycogen synthase kinase-3β. Journal of Biomedical Science, 21(1), 38.

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Palmitate-induced Inflammatory Signaling Pathways

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A total of 50 mM palmitate (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) was dissolved in 1 ml 99% ethanol and mixed with 9 ml 10% bovine serum albumin (BSA; Sigma-Aldrich; Merck KGaA), from which a 5 mM palmitate-10% BSA stock solution was prepared, as previously described (23 (link)). Antibodies against phosphorylated (p)-JNK (cat. no. 9255S), JNK (cat. no. 3708S), p-ERK1/2 (cat. no. 9106S) and ERK1/2 (cat. no. 4696S) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Antibodies against RIP140 (cat. no. ab42125) and β-actin (cat. no. sc-130300) were purchased from Abcam (Cambridge, MA, USA) and Santa Cruz Biotechnology, Inc. (Dallas, TX, USA), respectively. Goat anti-rabbit immunoglobulin (Ig)-G (heavy and light chain) secondary antibody conjugated to horseradish peroxidase (HRP; cat. no. BS13278) was obtained from Bioworld Technology, Inc. (St. Louis Park, MN, USA). ELISA kits for mouse TNF-α (cat. no. EK0527) and IL-6 (cat. no. EK0411) were purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China).

Zou R., Xue J., Huang Q., Dai Z, & Xu Y. (2017). Involvement of receptor-interacting protein 140 in palmitate-stimulated macrophage infiltration of pancreatic beta cells. Experimental and Therapeutic Medicine, 14(1), 483-494.

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Insulinoma INS-1 Cell Culture and FFAs

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The rat insulinoma INS-1 cell line kindly provided by Dr. Christopher B. Newgard (Duke University Medical Center, USA) was routinely passaged and cultured as described previously (28 (link)). INS-1 β-cells between passages 20–30 were used. Each cell experiment was performed in triplicates with different passage of cells. For FFA exposure, palmitate (Sigma-Aldrich, USA) was dissolved in 99% ethanol to a concentration of 100 mM, and then mixed with 10% BSA in serum-free DMEM to make a 5 mM palmitate stock solution and used at a final concentration of 0.5 mM. MnTMPyP (MerckSharp & Dohme, USA) was diluted in water. It was added together with palmitate and maintained in the medium for 24 h after adenovirus transduction. To verify whether Plin5 was associated with PI3K/Akt or ERK pathways, LY294002 (10 μM, PI3K/Akt inhibitor, Millipore, USA) or U0126 (20 μM, ERK inhibitor, Calbiochem-Novabiochem, USA) was used 1 h before palmitate treatment and INS-1 cells treated with DMSO was used as the control.

Zhu Y., Ren C., Zhang M, & Zhong Y. (2020). Perilipin 5 Reduces Oxidative Damage Associated With Lipotoxicity by Activating the PI3K/ERK-Mediated Nrf2-ARE Signaling Pathway in INS-1 Pancreatic β-Cells. Frontiers in Endocrinology, 11, 166.

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Palmitate exposure on HUVECs

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HUVECs were purchased from the American Type Culture Collection (Manassas, VA, USA; PCS-100-010) and cultured in M199 medium (HyClone; GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 20% fetal bovine serum (FBS; HyClone; GE Healthcare Life Sciences) at 37°C in an atmosphere containing under 5% CO₂. Cells were passaged every 2–3 days once they reached maximum confluence. Cells were incubated in M199/10% FBS medium supplemented with 0.05, 0.1, 0.2, 0.4 or 0.6 mM palmitate (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) preconjugated with FFA-free bovine serum albumin (BSA; Sigma-Aldrich; Merck KGaA) at a 1:1 molar ratio. Control cells were grown with the same medium containing FFA-free BSA. If not stated otherwise, cells were incubated for 1 h with 10 µΜ cathepsin L inhibitor (z-FF-FMK; cat. no. 219421; Calbiochem; EMD Millipore, Billerica, MA, USA) and cathepsin S inhibitor (z-FL-COCHO.H₂O; cat. no. 219393; Calbiochem; EMD Millipore) at 37°C, which was followed by incubation with palmitate or FFA-free BSA for 24 h at 37°C.

Zhang J., Shan Y., Li Y., Luo X, & Shi H. (2017). Palmitate impairs angiogenesis via suppression of cathepsin activity. Molecular Medicine Reports, 15(6), 3644-3650.

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Preparation of Fatty Acid-BSA Solutions

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The fatty acids palmitate (sodium palmitate, Sigma-Aldrich, St. Louis, MO, USA) and oleate (Sigma Aldrich, St. Louis, MO, USA) were prepared by dissolving 100 mmol/L of palmitate or oleate in 0.1 M NaOH at 70 °C. Then, these solutions were diluted 1/20 in 10% BSA (fatty acid-free bovine albumin, Sigma Aldrich, St. Louis, MO, USA) at 55 °C and stirred for 10 min. These 10X stock solutions (5 mM of palmitate or oleate in BSA) were sterilized by ultrafiltration (Merck Millipore filter 0.22 µm), aliquoted and stored at −20 °C. Each aliquot was thawed only once. As a control, the vehicle containing NaOH alone with BSA was used.

Lillo Urzúa P., Núñez Murillo O., Castro-Sepúlveda M., Torres-Quintana M.A., Lladser Caldera Á., Quest A.F., Espinoza Robles C., Llanos Vidal P, & Wehinger S. (2020). Loss of Caveolin-1 Is Associated with a Decrease in Beta Cell Death in Mice on a High Fat Diet. International Journal of Molecular Sciences, 21(15), 5225.

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Modulating Insulin Resistance and Mitochondrial Function in C2C12 Myotubes

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C2C12 myoblasts (ATCC, Manassas, VA, USA) were maintained in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA). Two days after the cells reached confluence, differentiation was induced with 1×high-glucose DMEM with 2% horse serum (Invitrogen) for 4 days. To induce insulin resistance, C2C12 myotubes were treated with 500 µM palmitate (Sigma-Aldrich Co., St. Louis, MO, USA) for 24 hours. To induce mitochondrial dysfunction, C2C12 myotubes were treated with 10 µM antimycin A (Sigma-Aldrich Co.) for 48 hours. After pretreatment with antimycin A, palmitate, or dimethyl sulfoxide (DMSO), 100 µM Rg3 (Sigma-Aldrich Co.) or DMSO was added to the medium for 24 hours. In some experiments, 10 µM compound C (Sigma-Aldrich Co.) was added 1 hour before Rg3 treatment.

Kim M.J., Koo Y.D., Kim M., Lim S., Park Y.J., Chung S.S., Jang H.C, & Park K.S. (2016). Rg3 Improves Mitochondrial Function and the Expression of Key Genes Involved in Mitochondrial Biogenesis in C2C12 Myotubes. Diabetes & Metabolism Journal, 40(5), 406-413.

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