Blood was collected using lithium heparin (Deltalab, Barcelona, Spain) as an anticoagulant. Plasma was obtained by centrifugation (1500× g for 15 min at 4 °C) and stored at −80 °C in different aliquots until analysis. The samples to be analyzed for ghrelin were collected into a tube containing protease inhibitors (cOmplete ULTRA Protease Inhibitor Cocktail, Roche, NY, USA). The intestine and other tissues were carefully removed, measured and weighed. Different intestinal segments were excised from the duodenum, the ileum and the proximal colon. A sample of each segment was immediately frozen in liquid nitrogen and stored for mRNA analysis. Another piece of the duodenum, ileum and proximal colon were taken for the ex vivo study. The cecum was quickly weighed before and after the removal of cecal content. The cecal content was immediately frozen in liquid nitrogen and stored. All samples were stored at −80 °C until analysis.
Lithium heparin
Manufactured by Deltalab
Sourced in Spain
Lithium heparin is an anticoagulant used in blood collection tubes to prevent blood clotting. It inhibits the coagulation cascade, allowing for the preservation of blood samples for analysis.
Automatically generated - may contain errors
2 protocols using lithium heparin
1
Isolation and Preservation of Biological Samples
2
Plasma Analytes and Gut Tissue Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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At the end of the study, the animals were fasted for 1-4 hours, anaesthetized with sodic pentobarbital (70 mg/kg body weight; Fagron Iberica, Barcelona, Spain) and exsanguinated from the abdominal aorta. The blood was collected using lithium heparin (Deltalab, Barcelona, Spain) as an anticoagulant. The blood was collected and aliquoted. Samples to measure GLP-1 were treated with a commercial Dipeptidyl peptidase-4 inhibitor (DPPIV, Millipore, Madrid, Spain) and a serine protease inhibitor (Pefabloc SC, Roche, Barcelona, Spain). The samples to be analysed for active ghrelin were treated with the serine protease inhibi nd 0.1 M HCl. All he mple we e ed -80ºC. Plasma was obtained by centrifugation (1500g, 15 minutes, 4°C) and stored at -80°C until analysis. The caecum was quickly weighed before and after caecal content removal. Intestinal segments were measured. The caecal content together with stomach and intestinal segments from the duodenum, jejunum, ileum and proximal colon were immediately frozen in liquid nitrogen and then stored at -80º C for further analysis.
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