Clotting activator serum

We previously evaluated sICAM (soluble intercellular adhesion molecule) [18 (link)]. Measurement was drawn from the same routine blood sample in S-Monovette 9 mL, Clotting Activator/Serum (Sarstedt, Nümbrecht, Germany, ref. 02.1063) and was allowed to clot for 30 min. After centrifugation at 2500× g for 10 min at room temperature, the supernatant was collected in aliquots and frozen at −80 °C until analysis. Soluble ICAM-1 levels were measured using the human ICAM-1/CD54 nonallele-specific Quantikine ELISA kit (Cat. no. SCIM00, R&D Systems, Inc., Minneapolis, MN, USA) according to the manufacturer’s instructions. Every blood sample was measured in duplicate.

Blood samples were collected by peripheral venous puncture the day before surgery during routine preoperative blood draw. Soluble ICAM-1 and CEA levels were determined in samples from the same blood draw. Blood was drawn in S-Monovette 9 ml, Clotting Activator/Serum (Sarstedt, Nümbrecht, Germany, ref. 02.1063) and was allowed to clot for 30 min. After centrifugation at 2500×g for 10 min at room temperature, the supernatant was collected in aliquots and frozen at − 80 °C until analysis. Soluble ICAM-1 was measured using the human ICAM-1/CD54 non-allele-specific Quantikine ELISA kit (Cat. no. SCIM00, R&D Systems, Inc. Minneapolis, USA) following the manufacturer’s instructions. Every blood sample was measured in duplicates. The mean value of sICAM-1 was calculated and used for further analysis.

Mice received 100 µL of Ovalbumin (OVA) solution (400 µg/mL) dissolved in sterile PBS one hour prior to the end of reperfusion. Afterwards blood was collected via puncture of the heart and drained into a micro tube (1.3 mL, Clotting Activator/Serum, SARSTEDT AG & Co., Nümbrecht, Germany). After 24 h storage at 4 °C, serum was obtained by centrifugation at 10,000 rpm for 5 min at room temperature. OVA concentration in serum was determined with a commercially available enzyme-linked immunosorbent assay kit (General Ovalbumin (OVA) ELISA Kit, MyBioSource, San Diego, CA, USA) according to manufacturer’s instruction.