Osteonectin (ON) ELISA Assay Kit


The Osteonectin (ON) ELISA Assay Kit is an enzyme immunoassay intended for the quantitative determination of osteonectin in serum. For research use only. The Eagle Biosciences Osteonection (ON) ELISA Assay Kit is not for use in diagnostic procedures.

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Osteonectin ON ELISA Assay Kit

Osteonectin ON ELISA Assay Kit manufactured in Germany by Immundiagnostik

Size: 1×96 wells
Incubation Time: Overnight; 1 h; 10-20 min
Sample Type: Serum
Sample Size: 25 µl
Alternative Names: ON, SPARC, BM-40
For Research Use Only
Controls Included

Assay Principle

This Osteonectin ON ELISA Assay Kit is designed for the quantitative determination of osteonectin. The test principle is based on a competition between antigen in the sample or standards and the antigen coated on the wells of microplate. Standards or samples are transferred with the primary antibody against osteonectin directly into the precoated microtiter plate. The antigen in the samples competes with the antigen immobilised on the microtiter plate for the free binding site of the specific antibodies against osteonectin. A peroxidase-conjugated antibody is used for detection and quantification, and tetramethylbenzidine (TMB) as a peroxidase substrate. The enzymatic reaction is terminated by acidic stop solution. A dose response curve of absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from standard. The colour change is inversely proportional to the amount of analyte (sample or control). Osteonectin, present in the samples is determined directly from this curve.

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Additional Information

Assay Background

Osteonectin (SPARC, BM-40) is a 32,700 molecular weight, Ca2+-binding glyco-protein which is a synthetic product of osteoblasts, endothelial cells, and megakaryocytes. Osteonectin is involved in tissue remodeling both from the perspective of bone metabolism and endothelial cell proliferation and repair. Therefore, the quantitative analysis of this protein could serve as a useful marker for the researcher studying the biochemical processes.

Storage and Preparation of Reagents

• To run the assay more than once, ensure that reagents are stored at the conditions stated on the label. Prepare only the appropriate amount necessary for each run. The kit can be used up to 2 times within the expiry date stated on the label.
• Preparation of the wash buffer: The wash buffer concentrate (WASHBUF) has to be diluted with ultrapure water 1:10 before use (100ml WASHBUF + 900ml ultrapure water), mix well. Crystals could occur due to high salt concentration in the concentrate. Before dilution, the crystals have to be redissolved at room temperature or in a water bath at 37°C. The WASHBUF is stable at 2–8°C until the expiry date stated on the label. Wash buffer (1:10 diluted WASHBUF) can be stored in a closed flask at 2–8°C for 1 month.
• The standards (STD), controls (CTRL) and antibody (AB) can be stored at 2–8°C for 2 weeks. For a storage for more than two weeks the components have to be stored at -20°.
• All other test reagents are ready-to-use. Test reagents are stable until the expiry date (see label) when stored at 2–8°C.

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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