Rabbit ctx 2 elisa kit

Synovial fluid was collected from the operated knee before euthanization and centrifuged at 12,500 rpm for 10 min at 4 °C. Synovial concentrations of COMP, CTX-II, MMP-3, IL-1β, and TNF-α were measured using ELISAs in accordance with previous reports [15 (link),16 (link),49 (link),58 (link)], using methods similar to those described for the serum measurements. A rabbit COMP ELISA kit (MBS721182, MyBioSource, San Diego, CA, USA), rabbit CTX-II ELISA kit (MBS705896, MyBioSource, San Diego, CA, USA), rabbit MMP-3 ELISA kit (MBS2502146, MyBioSource, San Diego, CA, USA), rabbit IL-1β ELISA kit (LS-F23299, LSBio Seattle, WA, USA) and rabbit TNF-α ELISA kit (MBS2500169, MyBioSource, San Diego, CA, USA) were used.

Approximately 10 mL of whole blood was collected from the vena cava at euthanization, and the serum was separated by centrifugation at 3000 rpm for 10 min. Serum concentrations of COMP, CTX-II, and MMP-3 were measured using sandwich enzyme-linked immunosorbent assays (ELISAs) in accordance with previous reports [15 (link),16 (link)]. A rabbit COMP ELISA kit (MBS721182, MyBioSource, San Diego, CA, USA), rabbit CTX-II ELISA kit (MBS705896, MyBioSource, San Diego, CA, USA) and rabbit MMP-3 ELISA kit (MBS2502146, MyBioSource, San Diego, CA, USA) were used in this measurement. All ELISA procedures were performed according to the manufacturer’s instructions, except for the dilution ratio. Standards and samples (100 μL) were added to the appropriate wells and incubated for 2 h at 37 °C. The wells were aspirated, and a 1× biotin-labeled antibody (100 μL) was added to each well, followed by incubation for 1 h at 37 °C. The wells were then aspirated again, washed three times, and incubated with 1× horseradish peroxidase-conjugated avidin (100 μL) for 1 h at 37 °C. 3,3′,5,5′-Tetramethylbenzidine substrate (90 μL) was added to each well, followed by incubation for 15–30 min at 37 °C. Stop solution (50 μL) was then added to each well, and absorbance was measured at 450 nm using a microplate reader (Tecan, Männedorf, Switzerland).