Phosphatidylinositol

Manufactured by Avanti Polar Lipids

Sourced in United States

Phosphatidylinositol is a type of phospholipid found in the cell membranes of eukaryotic organisms. It plays a crucial role in various cellular processes, including signal transduction, membrane trafficking, and cytoskeleton organization.

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24 protocols using phosphatidylinositol

Lipidomic Analysis of Spirulina

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Lipid analysis of dry spirulina was performed by Avanti Polar Lipids (Alabaster, AL, USA). Modified Bligh and Dyer extractions (Bligh and Dyer 1959) were performed with EquiSPLASH™ Lipidomix™, and Lipidyzer™ standards (Avanti Polar Lipids), and different chain lengths of phosphatidylglycerol, phosphatidylserine, and phosphatidylinositol (Avanti Polar Lipids) were added as internal standards (IS). Total lipids were dried then resolvated in 1 mL of 1:1 methylene chloride:methanol. Samples were injected in triplicate without dilution for LC-MS analysis. The methodology used was hydrophophilic interaction liquid chromatography (HILIC), which separates lipids into classes and subclasses that span a narrow retention time window (Hines et al. 2017) . A standard of 18:1 cholesterol ester was used as an IS for stigmasterols and brassicasterols. Data are presented as ng/mg of sample. Values were calculated using a point calibration determined by multiplying area ratio averages for the analyte to IS and multiplying by the concentration of IS.

Ricigliano V., & Simone-Finstrom M. (2020). Nutritional and prebiotic efficacy of the microalga Arthrospira platensis (spirulina) in honey bees. Apidologie, 51(5), 898-910.

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Lipid Standards for Lipidomics

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The lipid standards phosphatidic acid (PA) 16:0/18:1, phosphatidylcholine (PC) 16:0/18:1, phosphatidylethanolamine (PE) 16:0/18:2, phosphatidylinositol (PI) 18:1/16:0, phosphatidylserine (PS) 16:0/18:1 were purchased from Avanti Polar Lipids, Inc. (Alabaster, Alabama, USA). Further reagent and material details are provided in the ESI. †

Rampler E., Coman C., Hermann G., Sickmann A., Ahrends R, & Koellensperger G. (2017). LILY-lipidome isotope labeling of yeast: in vivo synthesis of ¹³C labeled reference lipids for quantification by mass spectrometry. The Analyst, 142(11).

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Profiling Lipid Standards for Mass Spectrometry

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Cell culture reagents, Lipofectamine LTX and blasticidin-S were from Life Technologies. All HPLC solvents were MS grade (Carlo Erba Reagents, Italia). Phospholipid standards: C13:0 lysophosphatidylcholines (lysoPC); C25:0 phosphatidylcholines (PC); C12:0 sphingomyelin (SM); 12:0–13:0 phosphatidylserine (PS); 12:0–13:0 phosphatidylinositol (PI); 12:0–13:0 phosphatidylglycerol (PG); 12:0–13:0 phosphatidic acid (PA); 12:0–13:0 phosphatidylethanolamine (PE); C12 ceramide (Cer); glucosyl (β) C12 ceramide (GCer); lactosyl (β) C12 ceramide (LacCer); C17 mono-sulfo galactosyl(β) ceramide (D18:1/17:0) were purchased from Avanti Polar Lipids. All other reagents were from Sigma-Aldrich.

Buratta S., Urbanelli L., Sagini K., Giovagnoli S., Caponi S., Fioretto D., Mitro N., Caruso D, & Emiliani C. (2017). Extracellular vesicles released by fibroblasts undergoing H-Ras induced senescence show changes in lipid profile. PLoS ONE, 12(11), e0188840.

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Lipid Standards for Mass Spectrometry

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Acetonitrile, methanol, and 2-propanol (Optima LC-MS grade) were purchased from Fisher Chemical(Waltham, USA); methyl-tert-butyl ether (MTBE), ammonium formate, ammonium hydrogen carbonate (BioUltra grade) and chloroform (analytical reagent grade) were purchased from Sigma-Aldrich (St. Louis, USA) Deionized water was obtained from an in-house water purification system. The following standards, lyso phosphatidylethanolamine (LPE) 14:0, Lyso phosphatidylcholine (LPC) 13:0, phosphatidylcholine (PC) 13:0_13:0, phosphatidylethanolamine (PE) 14:0_14:0, phosphatidylserine (PS) 14:0_14:0, phosphatidylinositol (PI) 12:0–13:0, ceramide d18:1/17:0, glucosyl(ß) C8 ceramide, hexosyl1ceramide d18:1/8:0 and 06:0 sphingomyelin (SM) were purchased from Avanti Polar Lipids (Alabaster, USA). GM3 d18:1/18:0 d3 was purchased from Matreya LLC (State College, USA). Diacylglycerol (DG) 15:0_15:0 was purchased from Santa Cruz Biotechnology, Inc. (Dallas, USA). Glyceryl triheptadecanoate (TAG) 17:0_17:0_17:0 was purchased from Sigma Aldrich (St. Louis, USA) and glyceryl trihexadecanoate (TAG) 48:0 d5 was purchased from CDN Isotopes (Pointe Claire, Canada)

Selvalatchmanan J., Rukmini A.V., Ji S., Triebl A., Gao L., Bendt A.K., Wenk M.R., Gooley J.J, & Torta F. (2021). Variability of Lipids in Human Milk. Metabolites, 11(2), 104.

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Authentic Internal Standards for Lipidomic Analysis

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The authentic internal standards (IS) for lipidomic analysis, namely, phosphatidylcholine (PC) 13:0/13:0, phosphatidylethanolamine (PE) 15:0/15:0, phosphatidylinositol (PI) 8:0/8:0, lysophosphatidylcholine (LPC) 15:0, lysophosphatidylethanolamine (LPE) 13:0, and lysophosphatidylinositol (LPI) 13:0, were purchased from Avanti Polar Lipids (Alabaster, AL), while triacylglycerol (TG) 11:0/11:0/11:0 and free fatty acid (FFA) 17:0 were obtained from Sigma-Aldrich (St. Louis, MO). Unless specified, other chemicals and reagents were of the highest grade available and purchased from Sigma-Aldrich.

Chen Z., Sai S., Nagumo K., Wu Y., Chiba H, & Hui S.P. (2023). Distinctive serum lipidomic profile of IVIG-resistant Kawasaki disease children before and after treatment. PLOS ONE, 18(3), e0283710.

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Lipid Profiling Internal Standards

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For the authentic internal standards (IS), triacylglycerol (TG) 11:0/11:0/11:0 and free fatty acid (FFA) 17:0 were obtained from Sigma-Aldrich (St. Louis, MO, USA), and phosphatidylcholine (PC) 13:0/13:0, phosphatidylethanolamine (PE) 15:0/15:0, phosphatidylinositol (PI) 8:0/8:0, lysophosphatidylcholine (LPC) 15:0, lysophosphatidylethanolamine (LPE) 13:0, and lysophosphatidylinositol (LPI) 13:0 were purchased from Avanti Polar Lipids (Alabaster, AL, USA). Cardiolipin (CL) 15:0/15:0/15:0/15:0 was synthesized in our laboratory previously [16 (link)]. Other chemicals and reagents, which were of the highest grade available, were purchased from Sigma-Aldrich unless specified. The mixed solution of all the IS was prepared with methanol (containing 0.006% butylated hydroxytoluene, w/v) and stored at −80 °C for no more than two weeks before use.

Wu Y., Chen Z., Fuda H., Tsukui T., Wu X., Shen N., Saito N., Chiba H, & Hui S.P. (2021). Oxidative Stress Linked Organ Lipid Hydroperoxidation and Dysregulation in Mouse Model of Nonalcoholic Steatohepatitis: Revealed by Lipidomic Profiling of Liver and Kidney. Antioxidants, 10(10), 1602.

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Comprehensive Lipidomics Analytical Standards

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HPLC-grade acetonitrile (ACN), methanol (MeOH), and isopropanol (IPA) were purchased from Thermo Fisher Scientific, U.S.A. HPLC-grade methyl-tert-butyl ether (MTBE), ammonium formate, and formic acid were purchased from Sigma–Aldrich (St. Louis, MO, U.S.A.). Ultrapure water was obtained by a Milli-Q system (Millipore, Billerica, MA). SPLASH internal standards (330707, SPLASH™ Lipidomix Mass Spec Standards) were purchased from Avanti Polar Lipids (Alabaster, U.S.A.), including lyso-phosphatidylcholine (LPC) 18:1 (d7), 25 μg/ml; lyso-phosphatidylethanolamine (LPE) 18:1 (d7), 5 μg/ml; phosphatidylcholine (PC) 15:0-18:1 (d7), 160 μg/ml; phosphatidylethanolamine (PE) 15:0-18:1 (d7), 5 μg/ml; phosphatidylglycerol (PG) 15:0-18:1 (d7), 30 μg/ml; phosphatidylserine (PS) 15:0-18:1(d7), 5 μg/ml; phosphatidylinositol (PI) 15:0-18:1 (d7), 10 μg/ml; phosphatidic acid (PA) 15:0-18:1 (d7), 7 μg/ml; sphingomyelin (SM) d18:1-18:1 (d9), 30 μg/ml; cholesterol (d7), 100 μg/ml; ceramide (Cer) 18:1 (d7), 350 μg/ml; monoglyceride (MG) 18:1 (d7), 2 μg/ml; diglyceride (DG) 15:0-18:1 (d7), 10 μg/ml; and triglyceride (TG) 15:0-18:1 (d7)-15:0, 55 μg/ml

Bai Y., Huang W., Li Y., Lai C., Huang S., Wang G., He Y., Hu L, & Chen C. (2021). Lipidomic alteration of plasma in cured COVID-19 patients using ultra high-performance liquid chromatography with high-resolution mass spectrometry. Bioscience Reports, 41(3), BSR20204305.

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Peptide Synthesis and Antifungal Assay

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For peptide synthesis, fluoren-9-ylmethoxycarbonyl (FMOC) amino acids, rink amide 4-methylbenzhydrylamine resin and other chemicals were purchased from Calbiochem-Novabiochem (La Jolla, CA, USA). Acrylamide, ergosterol and MTT were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cholesterol (CH, from porcine liver), L-α-phosphatidylethanolamine (PE, from Escherichia coli), egg yolk L-α-phosphatidylcholine (PC), L-α-phosphatidylglycerol (PG, from E. coli), sphingomyelin (SM) and phosphatidylinositol (PI) were obtained from Avanti Polar Lipids (Alabaster, AL, USA). Calcein, 5-carboxytetramethylrhodamine (TAMRA), 3,3′-diethylthiodicarbocyanineiodide (DiSC₃-5) and SYTOX Green and Propidium iodide (PI) were acquired from Molecular Probes (Eugene, OR, USA). All other reagents purchased were analytical grade. Double distilled water was used in the preparation of buffers (Millipore Co., Bedford, MA, USA). The fungal strains Candida albicans (C. albicans), Candida catenulate (C. catenulate), Candida intermedia (C. intermidia), Candida rugosa (C. rugosa), Candida glabrata (C. glabrata) and Candida melibiosica (C. melibiosica) isolates were obtained from the Korean Collection for Type Cultures (KCTC). Drug resistant C. albicans (CCARM 14001, CCARM 14007 and CCARM 140020) were collected from the Culture Collection of Antibiotic-Resistant Microbes (CCARM).

Ramamourthy G., Park J., Seo C., J. Vogel H, & Park Y. (2020). Antifungal and Antibiofilm Activities and the Mechanism of Action of Repeating Lysine-Tryptophan Peptides against Candida albicans. Microorganisms, 8(5), 758.

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Lipid Standards for Biochemical Assays

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Diacylglycerol (DAG, #800515) was purchased from Sigma-Aldrich. Phosphatidylcholine (PC 34:1, #850475), phosphatidylinositol (PI 34:1, #850142), phosphatidylserine (PS 34:1, #840032), phosphatidylethanolamine (PE 34:1, #850757), phosphatidylglycerol (PG 36:1, #840503), phosphatidic acid (PA 34:1, #840857), cardiolipin (CL 72:4, #710335) were purchased from Avanti polar lipids.

van 't Klooster J.S., Cheng T.Y., Sikkema H.R., Jeucken A., Moody B, & Poolman B. (2020). Periprotein lipidomes of Saccharomyces cerevisiae provide a flexible environment for conformational changes of membrane proteins. eLife, 9, e57003.

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Lipidomic Analysis Protocol

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The LC/MS-grade isopropanol, methanol, and chloroform were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). The mobile-phase additive ammonium acetate was purchased from Sigma-Aldrich (St. Louis, MO). The internal standards such as, phosphatidylethanolamine (PE 15:0-15:0), phosphatidylinositol (PI 8:0-8:0), phosphatidylcholine (PC 13:0-13:0), lysophosphatidylcholine (LPC) 15:0, lysophosphatidylethanolamine (LPE) 13:0, and lysophosphatidylinositol (LPI) 13:0, were obtained from Avanti Polar Lipids (Alabaster, AL). Triacylglyceride (TAG) 11:0-11:0-11:0 and fatty acid 17:0 were obtained from Sigma-Aldrich (St. Louis, MO). TK-16 (Dextran) from Mastutani Chemical Industry Co., Ltd. (Hyogo, Japan), NZMP acid casein from Fonterra Cooperative Group Ltd. (Auckland, New Zealand), sucrose from Nippon Beet Sugar Manufacturing Co., Ltd. (Tokyo, Japan), soybean oil from J-Oil Mills, Inc. (Tokyo, Japan), crystalline cellulose from Ceolus PH-102, Asahi Kasei Chemicals Corp., (Tokyo, Japan), AIN93G mineral mix from MP Biomedicals (USA), AIN-93VX vitamin mix from CLEA Japan Inc. (Tokyo, Japan), choline bitartrate from Wako Pure Chemical Industries Ltd. (Osaka, Japan), lard from Bell Shokuhin Co., Ltd. (Sapporo, Japan) and all other reagents of analytical grade were purchased from Kanto Chemical Co., Inc. (Tokyo, Japan) unless otherwise specified.

Gowda S.G.B., Gao Z.J., Chen Z., Abe T., Hori S., Fukiya S., Ishizuka S., Yokota A., Chiba H, & Hui S.P. (2020). Untargeted Lipidomic Analysis of Plasma from High-fat Diet-induced Obese Rats Using UHPLC-Linear Trap Quadrupole-Orbitrap MS. Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 36(7).

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