The assay is based on the simultaneous binding of the collagen fragments by two monoclonal antibodies, one of which will subsequently mediate the binding to the solid surface of the microtitre plate. Standards, control, or unknown samples are pipetted into the appropriate well of a microtitre plate coated with streptavidin, and subsequently a mixture of a biotinylated antibody and a peroxidase-conjugated antibody is added. Then a complex between the collagen fragments and the two antibodies is generated, and the complex will bind to the streptavidin surface via the biotinylated antibody. Following this one step incubation at room temperature the wells are washed and a chromogenic substrate added. The colour reation is stopped with sulphuric acid and the absorbance is measured.
Size
96 wells
Concentration
n/a
Applications
colorimetric ELISA or EIA
Other Names
crosslaps, fragments of type I collagen released into bone cell culture supernatants during bone resorption in vitro kit, assay, immunoassay