Vacuette serum clot activator tube

For investigation of pre-, peri- and postoperative stress levels and a possible influence of the chosen opioid, solid phase, competitive, chemiluminescent enzyme immunoassay (LKCO1, ImmuliteTM 1000, Siemens Diagnostics, Malvern, PA, USA) was performed to assess the serum cortisol concentrations. For this purpose, blood was taken at 3 hourly intervals preoperatively, as well as 5 min after initiation of the surgery. Additional samples were taken at 15 min intervals between 30 and 120 min., as well as 150 and 180 min. and 6 h postoperatively. All samples were drawn into VACUETTE^® Serum Clot Activator Tubes, Greiner Bio-One GmbH, Kremsmuenster, Austria).

On d 50, blood was collected from 2 birds per pen (16 per treatment) from the wing vein using a 21-gauge needle. It was placed into purple-top EDTA-coated vacutainer tubes and analysed for haematology using an automated IDEXX LaserCyte Hematology Analyzer (IDEXX Laboratories (Pty) Ltd., Johannesburg, South Africa), whilst blood collected into red top Vacuette® Serum Clot Activator tubes without EDTA (Greiner Bio-One, GmbH, Frickenhausen, Germany) was analysed for biochemistry using an automated IDEXX Vet Test Chemistry Analyzer (IDEXX Laboratories (Pty) Ltd., Johannesburg, South Africa).

IgG3 was purified from six serum samples (average donor age: 44.5 ± 9 years; three male and three female; details are listed in supplemental Table S1). The serum samples were collected from healthy donors with informed consent in compliance with the institutional ethical board. Venous blood was collected in a 9-ml Vacuette serum clot activator tube (Greiner BioOne, Kremsmünster, Austria) and incubated at room temperature for 30 min, followed by centrifugation for 15 min at 1800 g. The serum fraction was then collected and stored at −20 °C. IgG was isolated by running the serum over a HiTrap Protein G HP column (GE Healthcare, Buckinghamshire, UK) and eluted with 0.1 m glycine-HCl, pH 2.7. The eluate containing IgG was then applied to a HiTrap MabSelect SuRE column packed with recombinant Protein A (GE Healthcare), and the IgG3-containing flow-through was concentrated using an Amicon Ultra-15 centrifugal filter device 10 kDa (Merck Millipore, Darmstadt, Germany) and dialyzed against PBS using a Slide-A-Lizer Dialysis Cassette, 10K MWCO (Dionex/Thermo Scientific, Sunnyvale, CA).