Osteopontin Human (OPN) ELISA Kit

Documentation

Specifications

• 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
• Description

  Assay Principle: 

  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.

  Background:

  Osteopontin (OPN) is vital in mineralization, blood vessel formation, cell survival, inflammation, and inhibition of iNOS expression, which protects tumor cells from NO-mediated macrophage cytotoxic attack. It is also 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. In addition, OPN mRNA expression increases approximately 40-fold in infarct tissue after myocardial infarction.