The serum levels of procollagen type 1 amino-terminal propeptide (P1NP), sclerostin and type I collagen cross-linked c-terminal peptide (CTX-1) were measured using the serum from mice by the Rat/Mouse PINP EIA (Immunodiagnostic Systems Limited, AC-33F1), SOST/Sclerostin Quantikine ELISA Kit (R&D, MSST00) and RatLapsTM (CTX-I) EIA (Immunodiagnostic Systems Limited, AC-06F1) according to the manufacturer's instruction.
Rat mouse pinp eia
Manufactured by Immunodiagnostic Systems
Sourced in United Kingdom
The Rat/Mouse PINP EIA is a quantitative enzyme immunoassay designed for the in vitro measurement of procollagen type I N-terminal propeptide (PINP) in rat and mouse serum or plasma samples. The assay provides a reliable tool for the assessment of bone formation activity in rodent studies.
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4 protocols using rat mouse pinp eia
1
Serum Markers of Bone Metabolism
2
Serum Biomarkers of Bone Turnover
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Female mice were harvested at 5 weeks of age. Blood samples were collected immediately after euthanization by opening the peritoneal cavity and revealing the posterior vena cava for puncture using a 25G needle and a 1‐mL syringe. The blood samples were collected into Micro 1.1‐mL Z‐Gel tubes (Sarstedt, Numbrecht, Germany) and centrifuged at 10,000g for 10 minutes at room temperature to separate the serum. Serum levels for the N‐terminal propeptide of type I procollagen (PINP) and cross‐linked C‐terminal telopeptide of type I collagen (CTX‐I) were measured using commercial enzyme immunoassay (EIA) kits (Rat/Mouse PINP EIA and RatLaps [CTX‐I] EIA, respectively) (Immunodiagnostic Systems, Boldon, UK), according to the manufacturer's instructions. The mice were not fasted before CTX‐I measurements.
3
Serum Bone Marker Quantification
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Blood samples were collected 10-weeks post-ovariectomy and allowed to coagulate. The serum levels of the bone formation marker PINP and the bone resorption marker CTX-1 were then measured using commercial enzyme immunoassays (EIA), according to the manufacturers’ instructions: Rat/Mouse PINP EIA (Immunodiagnostic Systems, East Boldon, South Tyneside, UK; AC-33F1) and RatLapsTM EIA (Immunodiagnostic Systems; # AC-06F1), respectively.
4
Bone Turnover Markers and Cytokines in Ovariectomized Mice
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To determine the concentrations of bone turnover markers (procollagen type I N-terminal propeptide (PINP); OPG; C-terminal telopeptide (CTX); RANKL), blood was harvested at an age of 12 or 36 weeks and 8 weeks after OVX or sham operation in microvettes (Sarstedt AG & Co., Nümbrecht, Germany), centrifuged at 13,000 × g for 7 min for serum collection and stored at −80°C until further use. Bone turnover markers were determined using RatLaps™ CTX-I EIA, Rat/Mouse PINP EIA (both from Immunodiagnostic Systems GmbH, Frankfurt am Main, Germany), mouse RANKL ELISA Kit (TNFSF11), and mouse Osteoprotegerin ELISA Kit (TNFRSF11B) (both from Abcam, Cambridge, United Kingdom) according to manufacturers’ protocols. Using a customized mouse Multiplex Cytokine Kit (ProcartaPlex, eBiscience, Frankfurt, Germany), the CXCL-1, CXCL-10, IL-6, monocyte chemoattractant protein-1 (MCP-1), macrophage colony-stimulating factor (M-CSF), and vascular endothelial growth factor (VEGF) concentrations were determined in the OVX and Sham mice.
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