Microvette 200 z gel

Blood samples were collected from the vena saphena after animal warming for 5 min under a heating lamp. 200 μl of blood was collected into tubes including the clotting activator (Microvette 200 Z-Gel, Sarstedt Ag & Co.). The blood samples were collected before the inoculation of the cancer cells (at study day -3) and before sacrifice (at study day 56). The blood samples were processed into serum as instructed by the manufacturer. The serum samples were analyzed for TRACP5b (tartrate-resistant acid phosphatase 5b, MouseTRAP and BoneTRAP Assays), PINP (procollagen type I N-terminal propeptide, Rat/Mouse PINP EIA), and CTX-I (C-terminal telopeptide of type I collagen, RatLaps EIA, all from IDS Systems). The measurements were done according to the protocol provided by the manufacturer, and the plates were read with the VICTOR2™ Multilabel Counter (PerkinElmer).

For insulin release studies, measurement of serum insulin concentration during Intraperitoneal glucose tolerance test (ipGTT) was performed by blood collection at 0 and 15 min post injection using a Microvette 200 Z-Gel (Sarstedt, Hildesheim, Germany), followed by centrifugation for serum separation and analysis. Samples were stored at −80 °C before measurement for insulin levels. Serum insulin levels were measured with Ultrasensitive Mouse Insulin ELISA (Mercodia AB).