Human resistin was purchased from Shenondoah Biotechnology (Warwick, PA, USA). Human insulin Actrapid was provided by Novo Nordisk (Bagsvaerd, Denmark). Sodium palmitic acid, DHA and FFA-BSA were purchased from Sigma-Aldrich (St. Quentin Fallavier, France) and all cell culture reagents were purchased from Euromedex (Souffelweyersheim, France). All other chemicals were purchased from Sigma-Aldrich (St. Quentin Fallavier, France).
Ffa bsa
Manufactured by Merck Group
Sourced in United States
FFA-BSA is a laboratory product developed by Merck Group. It is a fatty acid-bound bovine serum albumin (BSA) solution that can be used as a standard or control in various analytical procedures. The core function of FFA-BSA is to provide a consistent and well-defined source of fatty acid-bound albumin for research and testing purposes.
Automatically generated - may contain errors
Lab products found in correlation
Actrapid human insulin, by Novo Nordisk (1 mentions)
Fatty acid free bovine serum albumin, by Merck Group (1 mentions)
Heparin, by Merck Group (1 mentions)
Trypan blue, by Merck Group (1 mentions)
Endothelial cell growth supplement (ecgs), by Merck Group (1 mentions)
Dextran t 70, by Merck Group (1 mentions)
Rat tail collagen 1, by Serva Electrophoresis (1 mentions)
Recombinant human vegf165, by Thermo Fisher Scientific (1 mentions)
Palmitic acid, by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Methylcellulose, by Merck Group (1 mentions)
5 protocols using ffa bsa
1
Fatty Acid Signaling in Cell Cultures
2
Preparation of Fatty Acid-BSA Conjugate
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All tissue culture reagents were from Euroclone SpA except ECGS and heparin (Sigma Aldrich). Charcoal, dextran T-70, MTT, trypan blue, methylcellulose (cat #M0512), E2, T3, DHT, sodium acetate, palmitic acid, and free fatty acid-bovine serum albumin (FFA-BSA) were from Sigma Aldrich; rat tail collagen I from Serva; recombinant human VEGF-165 from Peprotech. palmitic acid was used as a conjugate to FFA-BSA. Briefly, a 75 mM stock solution of palmitic acid dissolved in ethanol was diluted 1:10 with a 10% FFA-BSA solution, and incubated overnight at 37°C before every experiment.
3
Preparation of Palmitate-BSA Complex
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Briefly, 2 mM palmitate (PA; Cat.-No 489662-1G, Sigma-Aldrich, Steinheim, Germany) in 0.15 mM sodium chloride solution (NaCl, Carl Roth GmbH, Karlsruhe, Germany) was incubated at 70 °C for 60 min. Simultaneously, 0.45 mM fatty acid-free BSA (FFA-BSA; Cat.-No A8806-1G, Sigma-Aldrich, Missouri, USA) was incubated at 37 °C, for 60 min, in 0.15 mM sodium chloride and filter sterilized. The palmitate and BSA solutions were mixed 1:2 and incubated at 37 °C, for 60 min, leading to a molar ratio of palmitate:BSA of 4.4:1. For the ToF-SIMS experiments, deuterated palmitate (dPA; Cat.-No P0500-10G, Sigma-Aldrich, Steinheim, Germany) was used. The stock solutions were supplemented into the cultivation media in final concentrations of 0.045 mM BSA and 0.2 mM PA.
4
Preparation of Palmitate-BSA Conjugate
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Sodium palmitate at 70 mM was solubilized in water by heating it up to 70°C. BSA free fatty acids (FFA BSA) (Sigma Aldrich) was dissolved in PBS and warmed up to 37°C with continuous stirring. Solubilized palmitate was added to BSA at 37°C with continuous stirring (for a final concentration of 5 mM in 7% BSA). The conjugated palmitate-BSA was aliquoted and stored at −80°C [26 (link)].
5
Intravenous Injection of VPC32183
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VPC 32183 (S), (S)-Phosphoric acid mono-(2-octadec-9-enoylamino-3-[4-(pyridine-2ylmethoxy)-phenyl]-propyl) ester (Ammonium Salt) (857340; Avanti Polar Lipids, Alabaster, Alabama, USA) was dissolved in 3% free-fatty acid BSA (FFA-BSA; Sigma-Aldrich) in saline.
VPC32183 was diluted to a concentration of 5 µM and 100 µl were injected intravenous in the tail vein at the time-points described in the text. In non-treated control mice only vehicle injections were performed (3% FFA BSA in saline).
VPC32183 was diluted to a concentration of 5 µM and 100 µl were injected intravenous in the tail vein at the time-points described in the text. In non-treated control mice only vehicle injections were performed (3% FFA BSA in saline).
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