After sedation, blood was collected via cardiac puncture in tubes containing EDTA, and the inhibitors, protease (Pefabloc, Sigma) and DPP-IV (Millipore), at final concentrations of 2 μg/mL and 0.02 μL/mL, respectively. Centrifugation was at 1300g for 15 minutes and storage was at −80°C. The small intestine was removed and flushed with cold PBS. Sections of duodenum and jejunum (Supplemental Methods ), 1 cm each, were sampled and stored at −80°C in RNALater (Invitrogen, Thermo Fisher Scientific, Waltham, MA) for subsequent RNA extraction and gene expression analysis (Table S2 ). The right tibia was fixed in 10% neutral-buffered formalin for histomorphometry. The left femur was resected of muscle and placed in sterile ice-cold saline for flushing and cell culture and the right was wrapped in saline-soaked gauze and frozen for mechanical testing.
Dppiv
Manufactured by Merck Group
Sourced in Spain
DPPIV is a laboratory equipment product manufactured by Merck Group. It is a device used for the measurement and analysis of dipeptidyl peptidase-IV (DPP-IV) activity. DPP-IV is an enzyme involved in various physiological processes. The DPPIV product provides researchers and scientists with a tool to study and quantify DPP-IV activity in samples.
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Lab products found in correlation
Rnalater, by Thermo Fisher Scientific (1 mentions)
Pefabloc, by Merck Group (1 mentions)
Milliplex map mouse metabolic hormone magnetic bead panel, by Merck Group (1 mentions)
Aprotinin, by Roche (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
Lithium heparin, by Deltalab (1 mentions)
Pefabloc sc, by Roche (1 mentions)
Sodic pentobarbital, by Fagron (1 mentions)
3 protocols using dppiv
1
Multimodal Tissue Sampling and Analysis
2
Plasma Hormone Analysis in Fasted and Fed Mice
3
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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