Osteopontin Human (OPN) ELISA Kit

Assay Principle:

The DetectX® Osteopontin Human (OPN) ELISA Kit is designed for the quantitative measurement of human Osteopontin levels in EDTA plasma, urine, milk, and tissue culture media. This sandwich ELISA has a run time of 2.5 hours. Read the kit insert for detailed instructions before conducting the assay. The kit includes an Osteopontin standard to create an accurate standard curve.

Protocol Summary:

• Add standards or diluted samples to the transparent microtiter plate coated with mouse anti-human Osteopontin antibody. Incubate at room temperature for 1 hour.
• Introduce the peroxidase-conjugated human Osteopontin monoclonal antibody.
• Incubate the mixture covered at room temperature for another hour, allowing the immunological reaction between the antibody and Osteopontin antigen in the sample.
• After incubation, wash away excess antibody. Then add the TMB substrate, which reacts with the bound antibody to generate a detectable signal.
• Use a plate reader to measure the signal at 450nm and calculate Osteopontin concentration using the standard curve.

Background:

Osteopontin (OPN) is a multifunctional protein, present in both secreted and intracellular forms, that interacts with several receptors and influences diverse signaling pathways. It plays significant roles in processes like mineralization, blood vessel formation, cell survival, and inflammation. OPN also contributes to tumor cell survival by inhibiting iNOS expression, thereby protecting tumor cells from nitric oxide-mediated macrophage cytotoxic attack.

OPN is an important molecule in neoplastic transformation and the development of some tumors, influencing invasiveness, progression, and metastasis. Elevated plasma OPN levels are indicative of colon and lung cancers, as well as metastatic carcinomas. It is also a strong marker for inflammatory diseases including Crohn’s disease, Lupus, and Rheumatoid Arthritis.

Moreover, OPN mRNA expression significantly increases in necrotic tissue following myocardial infarction. The DetectX® Osteopontin Human (OPN) ELISA Kit is an easy to use and efficient tool for researchers studying cancer biology, inflammatory diseases.