Isopropyl β thiogalactopyranoside iptg

All chemicals used for this study were obtained from the following sources: acetic acid from Carl Roth GmbH (Karlsruhe, Germany); sodium borohydride from Life Technologies, Inc. (Eggenstein, Germany); antibiotics and isopropyl-β-thiogalactopyranoside (IPTG) from Carl Roth GmbH (Karlsruhe, Germany), restriction enzymes from Thermo Fisher Scientific-Fermentas (Schwerte, Germany); the E. coli strain BL21 (DE3) from Invitrogen (Carlsbad, USA) and E. coli strain XL-1 from Stratagene (La Jolla, USA). Oligonucleotide synthesis was performed at BioTez Berlin Buch GmbH (Berlin, Germany). Nucleic acid sequencing was carried out at Eurofins MWG Operon (Ebersberg, Germany). HPLC grade methanol, chloroform, and water were from Fisher Scientific. 1,2-dipentadecanoyl-sn-glycero-3-phosphocholine (PC(15:0/15:0)) and 1,2-dipentadecanoyl-sn-glycero-3-phosphoethanolamine (PC(15:0/15:0)) from Avanti Polar Lipids (Alabaster, USA). All other reagents were from Sigma-Aldrich, unless otherwise stated. Human lung epithelial cells (A549) were purchased from ATCC and cultured according to the recommendations of the vendor.

The chemicals used for the different experiments were obtained from the following sources: Phosphate buffered saline without calcium and magnesium (PBS) from PAN Biotech (Aidenbach, Germany); nitrocellulose blotting membrane from Serva Electrophoresis GmbH (Heidelberg, Germany); EDTA (Merck KG, Darmstadt, Germany), arachidonic acid (AA) and authentic HPLC standards of HETE-isomers (15S-HETE, 15S/R-HETE, 12S/R-HETE, 12S-HETE, 8S/R-HETE, 5S-HETE) from Cayman Chem (distributed by Biomol GmbH, Hamburg, Germany); acetic acid from Carl Roth GmbH (Karlsruhe, Germany); sodium borohydride from Life Technologies, Inc. (Eggenstein, Germany); isopropyl-β-thiogalactopyranoside (IPTG) from Carl Roth GmbH (Karlsruhe, Germany); restriction enzymes from ThermoFisher (Schwerte, Germany); the E. coli strain Rosetta2 DE3 pLysS from Novagen (Merck-Millipore, Darmstadt, Germany); HEK293 cells from the German Collection of Microorganisms and Cell Culture GmbH (DSMZ, Braunschweig, Germany). The Bac-to-Bac∗ baculovirus expression system was purchased from Invitrogen Life Technologies (ThermoFisher, Schwerte, Germany). Oligonucleotide synthesis was performed at BioTez Berlin Buch GmbH (Berlin, Germany). Nucleic acid sequencing was carried out at Eurofins MWG Operon (Ebersberg, Germany). HPLC grade methanol, acetonitrile, n-hexane, 2-propanol, and water were from Fisher Scientific (Waltham, MA, USA).

The chemicals used for this study were obtained from the following sources: Arachidonic acid (AA) and authentic HPLC standards of HETE-isomers (15S-HETE, 15S/R-HETE, 12S/R-HETE, 12S-HETE, 5S/R-HETE, 5S-HETE) from Cayman Chem (distributed by Biomol GmbH, Hamburg, Germany); acetic acid from Carl Roth GmbH (Karlsruhe, Germany); sodium borohydride from Life Technologies, Inc (Eggenstein, Germany); isopropyl-β-thiogalactopyranoside (IPTG) from Carl Roth GmbH (Karlsruhe, Germany); restriction enzymes from ThermoFisher (Schwerte, Germany); the E. coli strain Rosetta2 DE3 pLysS from Novagen (Merck-Millipore, Darmstadt, Germany); HEK293 cells from the German Collection of Microorganisms and Cell Culture GmbH (DSMZ, Braunschweig, Germany). The Bac-to-Bac^∗ baculovirus expression system was purchased from Invitrogen Life Technologies (ThermoFisher, Schwerte, Germany). Oligonucleotide synthesis was performed at BioTez Berlin Buch GmbH (Berlin, Germany). Nucleic acid sequencing was carried out at Eurofins MWG Operon (Ebersberg, Germany). HPLC grade methanol, acetonitrile, n-hexane, 2-propanol and water were from Fisher Scientific (New Hampshire, United States).