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- Assay Type Sandwich ELISA
- Sample Types EDTA Plasma, Urine, Milk, Tissue Culture Media
- Sensitivity 0.246 ng/mL
- Species Human
- Assay Duration 2.5 Hours
- Samples/Plate 40 in Duplicate
- Readout Colorimetric, 450 nm
- Standard Curve
The Osteopontin Human (OPN) ELISA Kit quantitatively measures human Osteopontin levels in EDTA plasma, urine, milk, and tissue culture media. The Osteopontin Human (OPN) ELISA Kit is a sandwich ELISA with a run time of 2.5 hours. Please read the complete kit insert for more information before performing this assay.
Use our provided Osteopontin standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our mouse anti-human Osteopontin antibody and incubate at room temperature for 1 hour. Add the peroxidase-conjugated human Osteopontin monoclonal antibody. Then incubate the mixture covered at room temperature for 1 hour. The immunological reaction occurs between the peroxidase-conjugated human Osteopontin monoclonal antibody and the Osteopontin antigen in the sample or standard.
After the 1-hour incubation, wash away the excess peroxidase-conjugated Osteopontin monoclonal antibody and add the TMB substrate. The TMB substrate reacts with the bound peroxidase-conjugated Osteopontin monoclonal antibody generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the Osteopontin concentration in the samples.
Osteopontin (OPN) is an essential protein in secreted and intracellular forms. OPN interacts with various receptors of diverse signaling pathways depending on the state of the protein. It has vital functions in mineralization, blood vessel formation, cell survival, and inflammation. It also plays a role in tumor cell survival by inhibiting iNOS expression, which protects tumor cells from NO-mediated macrophage cytotoxic attack. OPN is a key molecule in neoplastic transformation and cancer development in various tumors, playing a critical role in invasiveness, progression, and metastasis. Plasma OPN is a positive indicator of colon and lung cancers and metastatic carcinomas. It also strongly indicates inflammatory diseases such as Crohn’s disease, Lupus, and Rheumatoid Arthritis. In addition, OPN mRNA expression increases approximately 40-fold in infarct tissue after myocardial infarction.