P9417

Manufactured by Merck Group

Sourced in United States

P9417 is a laboratory equipment product offered by Merck Group. It is designed for general laboratory use. The core function of P9417 is to perform specific tasks within a controlled laboratory environment.

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Lab products found in correlation

7 protocols using p9417

Yeast growth under POA stress

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Yeast precultures were grown overnight in SCD. The following day, cultures were diluted to OD600 0.5, and different concentrations of POA (Sigma-Aldrich; P9417) were added for 2 d. Prior to plating, cells were diluted to an OD600 of 0.3, and 5 µl was spotted on YPD agar plates. Cells were imaged at 2 d growth unless otherwise indicated.

Hariri H., Speer N., Bowerman J., Rogers S., Fu G., Reetz E., Datta S., Feathers J.R., Ugrankar R., Nicastro D, & Henne W.M. (2019). Mdm1 maintains endoplasmic reticulum homeostasis by spatially regulating lipid droplet biogenesis. The Journal of Cell Biology, 218(4), 1319-1334.

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Yeast Genetic Manipulation and Protein Expression

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All yeast strains used in this study are detailed in Table S1. Yeast genetic manipulations were conducted using classical yeast knock-in/out protocols. For ectopic protein expression, genes were cloned into pBP73-C or -G vectors encoding either the GPD promoter or CPY promoter. All plasmids used in this study are detailed in Table S2. Yeast transformations were performed using the lithium acetate method. We used Ds-Red HDEL to label the ER. Strains were selected using antibiotics or dropout selection media. All chemicals used to make the yeast media were purchased from Sigma-Aldrich (succinic acid, sodium hydroxide, ammonium sulfate, yeast nitrogen base without amino acids, or ammonium sulfate). Yeast media was supplemented with a final concentration of 2% dextrose unless otherwise indicated. oleate (Sigma-Aldrich; O1008) and POA (Sigma-Aldrich; P9417) were added to the culture media as indicated (0.2% oleate is equivalent to 6.32 mM, and 0.2% POA is equivalent to 7.04 mM).

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CD36 Binding Assay for Lipid Molecules

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OxLig-1 was synthesized and identified by our research group according
to previous methods.^{33 (link),53 (link)} Oxlig-1, oleic acid (O1008, Sigma),
palmitoleic acid (P9417, Sigma), stearic acid (S4751, Sigma), and
palmitic acid (P0500, Sigma) were dissolved in anhydrous ethanol to
a final concentration of 50 μg/mL and coated into 96-well polystyrene
plates (50 μL/well) by ethanol evaporation. The plates were
blocked with PBS containing 1% gelatin for 1 h. Then, recombinant
human CD36 protein with His tag (50 μg/mL, 10752H08H50, Invitrogen),
anti-His antibody (1:1000, ZSGB-BIO, China), and HRP-labeled secondary
antibody (1:1000, ZSGB-BIO, China) was added and incubated for 1 h,
Finally, the color was developed with o-phenylenediamine
buffer containing H₂O₂ and terminated by 2 N
H₂SO₄. Absorbance was measured at 492 nm. In
each step, the wells were extensively washed with PBS containing 0.05%
Tween-20.

Fu C., Xiang M.L., Chen S., Dong G., Liu Z., Chen C.B., Liang J., Cao Y., Zhang M, & Liu Q. (2023). Molecular Drug Simulation and Experimental Validation of the CD36 Receptor Competitively Binding to Long-Chain Fatty Acids by 7-Ketocholesteryl-9-carboxynonanoate. ACS Omega, 8(31), 28277-28289.

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Preparation of Fatty Acid Solutions

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FFA solutions were prepared as previously described [36 (link)]. In summary, C16:0 (#P0500, Sigma, St Louis, USA), C16:1 (#P9417, Sigma, St Louis, USA), C18:0 (#S4751, Sigma, St Louis, USA), C18:1 (#O1008, Sigma, St Louis, USA) and C18:2 (#L1376, Sigma, St Louis, USA) were first dissolved in NaOH and then complexed with fatty acid free, low endotoxin BSA (#A8806, Sigma, St Louis, USA) at a FFA:BSA molar ratio of 3.4:1.

L’homme L., Sermikli B.P., Staels B., Piette J., Legrand-Poels S, & Dombrowicz D. (2020). Saturated Fatty Acids Promote GDF15 Expression in Human Macrophages through the PERK/eIF2/CHOP Signaling Pathway. Nutrients, 12(12), 3771.

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NEFA-Induced Dose-Dependent Effects on BGCs

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The NEFA stock (50 mM) solution contained oleic acid (21.75 mM, O1008, Sigma, St. Louis, MO, USA), linoleic acid (2.45 mM, L1376, Sigma, USA), palmitic acid (15.95 mM, P5585, Sigma, USA), stearic acid (7.2 mM, S4751, Sigma, USA), and palmitoleic acid (2.65 mM, P9417, Sigma, USA), which were dissolved with potassium hydroxide (0.1 M) at 60 °C, and the pH of the NEFA solution was adjusted to 7.4 using hydrochloric acid (1 M). To test the dose-dependent effect of NEFA in BGCs, the cells were seeded in 96-well plates and treated for different concentrations of NEFA (0, 0.3, 0.6, 0.9, 1.2, 1.5, 1.8, 2.1, and 2.4 mM) at 37 °C for 12 and 24 h. The optimum NEFA treatment concentration and processing time were selected and used for the subsequent experiment.

Lei Z., Ali I., Yang M., Yang C., Li Y, & Li L. (2023). Non-Esterified Fatty Acid-Induced Apoptosis in Bovine Granulosa Cells via ROS-Activated PI3K/AKT/FoxO1 Pathway. Antioxidants, 12(2), 434.

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Modulation of Type I Interferon Response by Fatty Acids

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To test the effects of free saturated and unsaturated fatty acids in modulating the type I interferon response, we prepared palmitate-BSA, palmitoleate-BSA, myristate-BSA, and sodium oleate-BSA conjugates. In brief, to prepare the BSA solution, 10% (w/v) fatty acid-free bovine albumin (A9418-5G, MilliporeSigma) was gradually added to ultrapure water at 52°C with gentle agitation until the BSA was fully dissolved. To prepare palmitate, palmitoleate, and myristate, palmitic acids (P0500, Sigma-Aldrich), palmitoleic acids (P9417, Sigma-Aldrich), and myristic acids (M3128, Sigma-Aldrich) were added to 0.1M NaOH (S2770, Sigma-Aldrich) and heated to 70°C until fully dissolved, yielding a concentration of 100 mM. Likewise, sodium oleate (O7501, Sigma-Aldrich) was added to ultrapure water at 55°C to yield a concentration of 100 mM. To conjugate fatty acids to BSA, the 100 mM stocks of palmitate, palmitoleate, myristate, and sodium oleate were added to 10% BSA at a ratio of 1:9 and heated at 55°C for 10 min. After conjugation, both the 10% (w/v) BSA solution and fatty acid-BSA solutions were filter-sterilized using a 0.45 μm PES filter (SLHVR33RS, Fisher Scientific), aliquoted, and stored at −20°C. Before use, solutions were warmed to 37°C. For final use, 200 μM of both fatty acid-BSA conjugates were used.

Heath B.R., Gong W., Taner H.F., Broses L., Okuyama K., Cheng W., Jin M., Fitzsimonds Z.R., Manousidaki A., Wu Y., Zhang S., Wen H., Chinn S.B., Bartee E., Xie Y., Moon J.J, & Lei Y.L. (2023). Saturated fatty acids dampen the immunogenicity of cancer by suppressing STING. Cell reports, 42(4), 112303.

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Preparation of Free Fatty Acid Solution

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Composition of FFA solution used in this study were based on blood FFA composition of dairy cows in periparturient period (Contreras et ). The stock FFA solution was prepared at 1 wk before the experiment and kept at -80°C. Briefly, the stock solution of FFA was prepared by diluting the FFA components in 0.1 M KOH at 60°C, and then the pH of the solution was adjusted to 7.4 with hydrochloric acid (1 M). The stock solution of FFA (52.7 mM) included oleic acid (22.9 mM; 01383, Sigma-Aldrich Co.), linoleic acid (2.6 mM; L1012, Sigma-Aldrich Co.), palmitic acid (16.8 mM; P55385, Sigma-Aldrich Co.), stearic acid (7.6 mM; S4751, Sigma-Aldrich Co.), and palmitoleic acid (2.8 mM; P9417, Sigma-Aldrich Co.). The stock solution of FFA was heated in a water bath (60°C) until transparent and then diluted in RPMI-1640 basic medium containing 2% low fatty acid BSA (B2064, Sigma-Aldrich Co.) to treat hepatocytes. Nuc (B20500; >98%, HPLC) was purchased from Shanghai Yuanye Biotechnology Co. Ltd. (Shanghai, China) and dissolved in 1 M hydrochloric acid, and then the pH was adjusted to 7.5 using sodium hydroxide. The stock Nuc solution used in this study was 295 mM.

Fang Z., Jiang X., Wang S., Tai W., Jiang Q., Loor J.J., Yu H., Hao X., Chen M., Shao Q., Song Y., Lei L., Liu G., Du X, & Li X. (2024). Nuciferine protects bovine hepatocytes against free fatty acid-induced oxidative damage by activating the transcription factor EB/peroxisome proliferator-activated receptor γ coactivator 1 alpha pathway. Journal of dairy science, 107(1).

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