Freezing extender used in this study was Tris-glucose based extender containing 2.4% (w/v) Tris (BDH, England), 1.4% (w/v) citric acid (BDH, England), 0.8% (w/v) glucose (Merck, Germany), 0.06% (w/v) sodium benzyl penicillin (M&H manufacturing co., Ltd., Samutprakarn, Thailand), 0.1% (w/v) streptomycin sulphate (M&H manufacturing co., Ltd., Samutprakarn, Thailand), and different concentrations of L-carnitine (Sigma-Aldrich, St. Louis, MO, USA) (0 mM L-carnitine, 12.5 mM, 25 mM, and 50 mM). This extender was further supplemented with 3% and 7% (v/v) glycerol for extender 1 and 2, respectively. Thawing medium used in this study was prepared similarly to freezing extender except that it did not contain egg yolk, L-carnitine or glycerol (Axner et al., 2004 (link)).
L carnitine
Manufactured by Merck Group
Sourced in United States, Italy, Poland
L-carnitine is a naturally occurring compound that plays a crucial role in the metabolism of fats. It is primarily responsible for the transport of long-chain fatty acids into the mitochondria, where they can be used for energy production. This lab equipment product is an important component in various biochemical and metabolic research applications.
Automatically generated - may contain errors
Lab products found in correlation
Taurine, by Merck Group (9 mentions)
Palmitic acid, by Merck Group (8 mentions)
Linoleic acid, by Merck Group (7 mentions)
Creatine, by Merck Group (6 mentions)
Arachidonic acid, by Merck Group (5 mentions)
Hydrocortisone, by Merck Group (5 mentions)
Glucose, by Merck Group (5 mentions)
Bovine serum albumin (bsa), by Merck Group (5 mentions)
L serine, by Merck Group (4 mentions)
Trypsin, by Merck Group (4 mentions)
Betaine, by Merck Group (4 mentions)
Triton x 100, by Merck Group (4 mentions)
Palmitoyl coa, by Merck Group (4 mentions)
Oleic acid, by Merck Group (4 mentions)
Creatinine, by Merck Group (4 mentions)
Formic acid, by Merck Group (4 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (4 mentions)
B27 supplement, by Thermo Fisher Scientific (4 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
108 protocols using l carnitine
1
L-Carnitine Supplemented Tris-Glucose Extender
2
Coated Blade Spray Mass Spectrometry
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The liquid chromatography-mass spectrometry (LC-MS) grade solvents (i.e., isopropanol, methanol, water, and acetonitrile) and additives (i.e., ammonium acetate, acetic acid, and formic acid) used in this work were purchased from Sigma Aldrich (Poznan, Poland). The probes were prepared using N, N-dimethylformamide ACS reagent and polyacrylonitrile. In addition, the phosphate-buffered saline used in the experiments was acquired from Sigma Aldrich (Poznan, Poland).
The standards selected for this work included Sphingosine (d17:1), LPE (17:1), PC (C16–18:1), PG (17:0–20:4), L-carnitine(trimethyl-d9), L-carnitine inner salt, phenylalanine, and phenylalanine(d8). The lipid short-hand notation presented herein was used in this manuscript.
Coated blade spray-mass spectrometry probes (CBS-MS) were prepared according to a procedure described elsewhere [18 (link)]. Two types of sorbents were used in the experiments: octadecyl (C18), which was purchased from Anchem (Toruń, Poland), and hydrophilic–lipophilic balance particles (HLB), which were manufactured and kindly provided by the University of Waterloo [32 (link)].
The standards selected for this work included Sphingosine (d17:1), LPE (17:1), PC (C16–18:1), PG (17:0–20:4), L-carnitine(trimethyl-d9), L-carnitine inner salt, phenylalanine, and phenylalanine(d8). The lipid short-hand notation presented herein was used in this manuscript.
Coated blade spray-mass spectrometry probes (CBS-MS) were prepared according to a procedure described elsewhere [18 (link)]. Two types of sorbents were used in the experiments: octadecyl (C18), which was purchased from Anchem (Toruń, Poland), and hydrophilic–lipophilic balance particles (HLB), which were manufactured and kindly provided by the University of Waterloo [32 (link)].
3
Bovine Embryo Culture and Tracking
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After insemination, oocytes were completely denuded from cumulus cells and spermatozoa by repeated pipetting with a glass pipette in the IVC medium, potassium
simplex optimized medium (KSOM) with amino acid (KSOMaa Evolve Bovine; Zenith Biotech, Bangkok, Thailand) supplemented with 5% NCS and 0.6 mg/ml L-Carnitine
(C0158, Sigma-Aldrich). Presumptive zygotes were subsequently washed thrice with IVC medium and cultured for 48 h in 100 µl droplets of IVC medium. Each droplet
contained approximately 20 presumptive zygotes. At 48 hpi, embryos with more than four cells were transferred from 35 mm dishes to well-of-the-well (WOW) dishes
(LinKID micro25, Dai Nippon Printing Co., Ltd., Tokyo, Japan) as previously described [10 (link)]. WOW dishes, which are 35 mm
in diameter, have 25 microwells (5 columns × 5 rows) and a circular wall in the center. A WOW dish can culture up to 25 embryos each with a single drop of
medium, and allows tracking of individual embryos. Pre-cultured IVC medium (100 µl) was placed within the circular wall and covered with mineral oil. At 168 to
180 hpi, embryos that had developed to or beyond the blastocyst stage were observed under an inverted microscope.
simplex optimized medium (KSOM) with amino acid (KSOMaa Evolve Bovine; Zenith Biotech, Bangkok, Thailand) supplemented with 5% NCS and 0.6 mg/ml L-Carnitine
(C0158, Sigma-Aldrich). Presumptive zygotes were subsequently washed thrice with IVC medium and cultured for 48 h in 100 µl droplets of IVC medium. Each droplet
contained approximately 20 presumptive zygotes. At 48 hpi, embryos with more than four cells were transferred from 35 mm dishes to well-of-the-well (WOW) dishes
(LinKID micro25, Dai Nippon Printing Co., Ltd., Tokyo, Japan) as previously described [10 (link)]. WOW dishes, which are 35 mm
in diameter, have 25 microwells (5 columns × 5 rows) and a circular wall in the center. A WOW dish can culture up to 25 embryos each with a single drop of
medium, and allows tracking of individual embryos. Pre-cultured IVC medium (100 µl) was placed within the circular wall and covered with mineral oil. At 168 to
180 hpi, embryos that had developed to or beyond the blastocyst stage were observed under an inverted microscope.
4
Cholesterol and Antioxidant Modulation by LC-SLNs
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L-Carnitine (LC; C7H15NO3 HCl) was purchased from Sigma-Aldrich (Sigma-Aldrich, ≥98% L-Carnitine, Cas. No. 6645-46-1; St. Louis, MO). L-Carnitine-loaded solid lipid nanoparticles (spray-dried LC-SLNs, with a content of ≥20% LC) were prepared and various physicochemical characterizations were evaluated (Eskandani et al., 2022 ). A serum cholesterol detection Assay kit was purchased from a Biosis (Athens, Greece). Yolk and bile cholesterol, total antioxidant capacity (TAOC ), total superoxide dismutase (TSOD ), and glutathione peroxidase (GSH-Px ) detection kit was purchased from a Randox (Antrium, UK). To assay the levels of glucose (GLU ), alanine aminotransferase (ALT ), aspartate aminotransferase (AST ), triglycerides (TG ), and high-density lipoprotein (HDL ), detection kits bought from Pars Azmoon (Tehran, Iran). Malondialdehyde (MDA ) was assessed by using an MDA kit that was purchased from Sigma-Aldrich (Poole, UK). For total mRNA extraction from the liver tissues, the TriReagent kit was bought from Sigma-Aldrich (St. Louis). All other materials not mentioned here were pharmaceutical grades and provided by Merck Co (Darmstadt, Germany).
5
LC-MS Metabolite Profiling Protocol
6
Optimized Cell Culture Media Formulations
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Tryptic soy broth (TSB), Luria broth (LB), D-glucose, phosphate-buffered saline (PBS), Dulbecco’s PBS (DPBS), Keratinocyte-SFM medium, DMEM (high glucose, GlutaMAX™ Supplement, pyruvate), and Ham’s F-12 Nutrient Mix were purchased from ThermoFisher Scientific (Waltham, MA). DermaLife K Keratinocyte Complete Medium with LifeFactors was obtained from Lifeline Cell Technology (Oceanside, CA). CnT-Prime 3D Barrier Medium was purchased from CELLnTEC Advanced Cell Systems AG (Zurich, Switzerland). Gentamicin, lysostaphin trimethoprim, sucrose, glycerol, mupirocin, neutral-buffered formalin solution (10%), and various supplements for skin culture media including hydrocortisone, isoproterenol, bovine insulin, selenious acid, L-serine, L-carnitine, bovine serum albumin (BSA), palmitic acid, linoleic acid, and arachidonic acid were obtained from Sigma-Aldrich (St. Louis, MI).
7
PFOA and l-carnitine Interactions
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PFOA, l -carnitine, and the chemicals used for ESR were purchased from Sigma (St. Louis, MO, USA). Fertile chicken eggs (Gallus gallus) were purchased from Linwen Trading Co. Ltd. (Jining, China). The antibody against p65 was purchased from Abcam (Cambridge, MA, USA). The antibody against GAPDH was purchased from ZSbio (Beijing, China). Other chemicals and consumables were of the highest grade obtainable.
8
Palmitate-Induced Lipid Metabolism Assay
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Cells pretreated as described above were incubated further for 16 h at 37°C in serum free M199 medium containing 0.30 mM palmitate (Sigma, USA) bound to 1% bovine serum albumin (BSA, Sigma, USA) and 0.05 mM L-carnitine (Sigma, USA). After that, the cells were washed, collected and diluted with phosphate-buffered saline (PBS, pH 7.2–7.4). Cellular TG and free fatty acid (FFA) concentrations were measured using Chicken Triglyceride (TG) and FFA ELISA Kits (Jiyinmei Biological Technology Co., LTD, Wuhan, China), according to the manufacturer's guidelines.
9
Antibody-based Mitochondrial Protein Analysis
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Monoclonal anti-actin antibody, Prussian Blue, pantothenate, pantethine, vitamin E, L-carnitine, thiamine, Luperox® and trypsin were purchased from Sigma Chemical Co. (St. Louis, MO). Anti-mitochondrial acyl carrier protein (mtACP), anti-NF-Y, anti-FOXN4 and anti-hnRNPA/B were purchased from Invitrogen/Molecular Probes (Eugene, OR). Anti-phospho-PGC1α was purchased from RD systems. NFS1 antibody and Omega 3 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-PANK2, anti-PGC1α, complex 1 activity kit and PDH activity kit were purchased from Abcam (Cambridge, UK). Anti-TFAM was purchased from Cell Signaling. BODIPY® 581/591 C11 was purchased from Thermo-Fisher (Waltham, MA). A cocktail of protease inhibitors (complete cocktail) was purchased from Boehringer Mannheim (Indianapolis, IN). The Immun Star HRP substrate kit was from Bio-Rad Laboratories Inc. (Hercules, CA).
10
Lipid Metabolism Assay Protocol
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L-carnitine and palmitate were from Sigma, and palmitoyl Coenzyme A ammonium salt from Avanti Polar Lipids, Inc.
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