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Specifications
- Assay duration:Multiple steps
- Assay Type (ELISA with LOV):Competitive
- Conjugate ELISA:Biotin
- Format:Pre-coated
- Host:
- Primary antibody reactivity:RatMouse
- Target protein:C-terminal telopeptide I, carboxy-terminal collagen I crosslinks
- Description:RatLaps™ ELISA (CTX-I)
- Size:1 kit
- Environmentally Preferable:
- Sample Type:Serum, urine, plasma, cell culture supernatant
- Detection Method:Colorimetric
- Shelf Life:See expiration date
- Detection Range:0 - 200 ng/ml
- Storage Temperature:2…8 °C
- Sample Volume:20 µl
- Sensitivity:2.0 ng/ml
- Regulatory Status:RUO
- Cat. No.:MSPP-AC06PL
Specifications
About this item
96 wells (1 kit) - ELISA is a competitive enzyme immunoassay for the quantitative measurement of CTX-I.
RatLaps™ (CTX-I) EIA is an enzyme-linked immunosorbent assay for the quantitative determination of bonerelated degradation products from C-terminal telopeptides of type I collagen in rat/mouse serum or urine and from rat/mouse bone released into cell culture supernatants by osteoclasts.
Type I collagen accounts for more than 90% of the organic matrix of bone and is synthesized primary in bone (1). During renewal of the skeleton bone matrix is degraded and consequently fragments of type I collagen is released into circulation. The resorption process can be studied in vitro by culturing bone cells on slices of bone or dentin. The RatLaps™ (CTX-I) are based on the observation that certain C-telopeptide degradation products from type I collagen released during osteoclastic bone resorption. With RatLaps™ (CTX-I) it is possible to measure this degradation products in rat/mouse serum and urine and bone cell culture supernatants (2-8).
The RatLaps™ (CTX-I) is based upon the competitive binding of a polyclonal antibody to soluble RatLaps antigens EKSQDGGR or to immobilized RatLaps antigens. Briefly, the polyclonal antibody is raised against a synthetic peptide having a sequence (EKSQDGGR) specific for a part of the C-terminal telopeptide α1 chain of rat type I collagen. For standardization of the RatLaps™ (CTX-I) a synthetic peptide (EKSQDGGR), which is specific for the C-terminal telopeptide α1 chain of type I collagen in rats has been used. During the pre-incubation step, biotinylated EKSQDGGR is immobilized by binding to the streptavidin-coated microtitre wells. The wells are emptied and washed. Standards, control, or unknown samples (culture supernatant or rat/mouse serum or urine) are pipette into appropriate wells, followed by a solution of a primary antibody (polyclonal rabbit). Following the primary-incubation step the wells are emptied and washed. In the secondary-incubation step a solution of a goat anti-rabbit antibody conjugated with peroxidase (secondary antibody) is added and binds to the polyclonal rabbit antibody. After the third washing step a chromogenic substrate (TMB) is added and the color reaction is stopped with sulfuric acid. Finally, the absorbence at 450 nm is measured with 650 nm as reference if possible. The absorbence level is inversely related to concentration of RatLaps antigens in the sample.