The polyclonal rabbit-anti-human CerS3 antibody was from Antikörper-online (Aachen, Germany; 1:50). The secondary antibodies were the swine anti-rabbit-FITC antibody (Dako, Hamburg, Germany; 1:30) and the donkey anti-rabbit-Alexa-Fluor 555 antibody (Thermo Fisher Scientific, Darmstadt, Germany). DAPI was from Sigma-Aldrich GmbH (Taufkirchen, Germany). The following specific inhibitors were used: PPARγ antagonist (GW9962, Enzo, Lörrach, Germany), PI3K inhibitor (LY294002; Sigma-Aldrich GmbH), ERK inhibitor (PD98059, New England Biolabs GmbH, Frankfurt, Germany), p38 MAPK inhibitor (SB203580, Sigma-Aldrich GmbH) and NFκB inhibitor (GIV 3727, Merck Millipore, Darmstadt, Germany). Methanolic sodium hydroxide solution, hexane standards, loganic acid, and loganin were from Carl Roth GmbH (Karlsruhe, Germany). Boron trifluoride–methanol complex was from Merck KGaA (Darmstadt, Germany), and standard methyl heptadecanoate was from Sigma-Aldrich GmbH. Orto-phosphoric acid, 85%, Ph. Eur. p.a. grade was supplied by VWR. Amarogentin and gentiopicroside were purchased from Chromadex Inc. (Santa Ana, CA, USA).
Boron trifluoride methanol complex
Manufactured by Merck Group
Sourced in Germany
Boron trifluoride-methanol complex is a chemical compound used as a laboratory reagent. It serves as a Lewis acid catalyst in organic synthesis reactions.
Automatically generated - may contain errors
Lab products found in correlation
Supelco 37 component fame mix, by Merck Group (2 mentions)
Pd98059, by New England Biolabs (1 mentions)
Sb203580, by Merck Group (1 mentions)
Swine anti rabbit fitc antibody, by Agilent Technologies (1 mentions)
Ortophosphoric acid, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Ly294002, by Merck Group (1 mentions)
Amarogentin, by ChromaDex (1 mentions)
Gentiopicroside, by ChromaDex (1 mentions)
Gc grade hexane, by Merck Group (1 mentions)
Hydranal titrant 5, by Merck Group (1 mentions)
Hydranal water standard 10, by Merck Group (1 mentions)
Hydranal solvent, by Merck Group (1 mentions)
Anhydrous sodium sulfate, by Merck Group (1 mentions)
5 protocols using boron trifluoride methanol complex
1
Immunohistochemical Analysis of CerS3 Expression
2
Fatty acid methyl ester analysis
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Sodium methoxide (0.5 N) was purchased from Sigma–Aldrich (St Louis, MO). Boron trifluoride methanol complex (35%) was obtained from Merck (Darmstadt, Germany). The FA methyl ester mixture of standards was supplied by Supelco (Supelco 37 Component FAME mix, Supelco, St. Louis, MO). All the other reagents were of ACS quality grade.
3
Lipid Extraction and Fatty Acid Profiling
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The phospholipid extracts were saponified with 0.5 N methanolic KOH and methylated with boron trifluoride-methanol complex (Merck, Darmstadt, Germany). The fatty acid methyl esters were separated by GC-MS on a column 60 m × 0.25 mm ID BPX70 × 0.25 µm.
4
Extraction and Characterization of Atlantic Salmon Oil
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Atlantic salmon (Salmo salar L.) was obtained from the local market (Timişoara, Romania) as a raw product in the spring of 2014. It was an aquaculture product of Norwegian origin. Only the meaty fish parts were used for oil extraction. GC-grade hexane (Sigma-Aldrich) was the main solvent used for raw and degraded fish oil dilutions. The Supelco 37 Component FAME mix (Sigma-Aldrich) and C8–C20 alkane standard solution (Fluka Chemie AG) were the main tools for identifying the FAMEs in derivatized raw and degraded fish oil. Anhydrous sodium sulfate (p.a., Merck & Co.) was used for drying the raw and degraded fish oil solutions. β-CD hydrate (>98%) from CycloLab (Budapest, Hungary) was used for ASO molecular encapsulation. Ethanol 96% (v/v, Chimopar, Bucharest, Romania) was used for β-CD complexation of ASO. A boron trifluoride–methanol complex (20%, Merck & Co., Inc.) was used for FA derivatization. Finally, Hydranal-Titrant 5, Hydranal-Solvent and Hydranal-Water Standard 10.0 (Sigma-Aldrich, Buchs, Switzerland) were used for the KFT water analysis of β-CD/ASO complexes.
5
Quantification of Phospholipase A2 Activity
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The phospholipid extracts were saponified with 0.5 N methanolic KOH and methylated with boron trifluoride-methanol complex (Merck). The fatty acid methyl esters were extracted with hexane and separated by gas chromatography on a capillary column coated with Supelcowax 10-bound phase 9 (i.d. 0.32 mm, length 30 m, film thickness 0.25 µm, Supelco, Bellafonte, PA, USA) fitted in a Perichrom gas chromatograph (Perichrom, Saulx-les-Chartreux, France). Phospholipase A2 activity was assayed by quantification of the fatty acid products by gas chromatography.
All the analyses for the determination of phospholipase A2 activity were performed in duplicate from different experiments.
All the analyses for the determination of phospholipase A2 activity were performed in duplicate from different experiments.
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