Elisa crosslaps kit

IgG, IgM, and IgA serum levels were measured using commercially available ELISA kits (Bethyl Laboratories, Nordic BioSite). Levels below the assay range were set to a level that was the lowest detectable level according to the kit.
As a marker for bone resorption, serum levels of C‐terminal type I collagen fragments (CTX‐I) were determined with a commercially available ELISA Crosslaps kit (Immunodiagnostic Systems, Boldon, UK). As a marker of bone formation, serum levels of N‐terminal propeptide of type I procollagen (P1NP) were assessed with a commercially available EIA P1NP kit (Immunodiagnostic Systems).
Levels of mBSA‐specific antibodies were assessed in serum of AIA mice using an in‐house ELISA, as described.⁽³³⁾ Briefly, ELISA plates were coated with 0.01 mg/mL mBSA, blocked and washed with casein (Sigma‐Aldrich) before adding the serum. Bound anti‐mBSA antibodies were detected by horseradish peroxidase (HRP)‐conjugated rabbit anti‐mouse IgG (DAKO, Agilent, Santa Clara, CA USA).

Blood was taken from the axillary vessels at termination, and serum was prepared using Multivette tubes (Sarstedt, Helsinborg, Sweden) standing in room temperature for 20 minutes followed by centrifugation. The serum samples were individually stored at –20°C. mBSA‐specific immune globulin G (IgG) was assessed in triplicate using ELISA as previously described.⁽²¹⁾ ELISA plates were coated with 0.01 mg/mL mBSA, blocked, and washed with milk powder (Carl Roth, Frankfurt, Germany) before adding serum. Bound anti‐mBSA antibodies were then detected by horseradish peroxidase (HRP)‐conjugated rabbit‐anti mouse IgG (Supplemental Table S1).
As a marker for osteoclast activity, serum levels of C‐terminal type I collagen fragments (CTX‐I) were determined with the ELISA Crosslaps Kit (Immunodiagnostic Systems, Boldon, UK).