Osteopontin N-Half ELISA Kit

INTENDED USE

Human osteopontin N-half ELISA kit is a reliable quantitative method for measuring OPN N-half (osteopontin N-half, N-terminal OPN) in human plasma, serum, tissue homogenates and other biological fluids. This assay has a sensitivity detection limit of 0.094 ng/ml.

BACKGROUND

Osteopontin (OPN) is a highly acidic calcium binding glycosylated phosphoprotein, also a secretory glycoprotein that was first isolated from the bone. It has a molecular weight which can vary as a result of phosphorylation and sugar chain formation, in general it can be anything from 44 to 66kDa. It has a unique characteristic in that it can assume a number of different molecular forms in vivo as a result of; phosphorylation, RNA splicing, degradation by protease, saccharification, sulfation and glycosylation. It also contains an Arg-Gly-Asp (RGD) amino acid sequence, this motif allows OPN to bind members of the integrin family (such as avb3) of cell surface receptors.

The following assay procedure is designed for measuring the N-terminal osteopontin (OPN N-half) fragment cleaved by thrombin. Therefore, any other OPN molecules that are not cleaved by thrombin will not be detected. It is known to be able to coexist with thrombin locally in tissues that are; inflamed, injured, vascularized and tumors. This ability to coexist with thrombin can help to increase the likelihood of proteolysis and there is growing evidence that this may have a vital physiological role.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti-OPN antibody.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

 TYPICAL RESULTS

For this OPN N-half ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.156, 0.312, 0.625, 1.25, 2.50, 5.0, 10.0 ng/ml.
Reactivity: Human
Sensitivity: 0.094 ng/ml
Range: 0.156 – 10 ng/ml
Principle: Sandwich
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for OPN N-half, no cross reactivity or interference between OPN N-half and analogues was detected.
– Recovery: Serum (88 – 98%), EDTA Plasma (86 – 102%), Heparin Plasma (85 – 102%).
– Linearity: Serum (86 – 104%), EDTA Plasma (85 – 101%), Heparin Plasma (86 – 99%).
– Precison Intra-Assay: CV < 5%.
– Precison Inter-Assay: CV < 5%.
– Stability: Less than 10%.

REFERENCES

1. Molecular cloning and sequencing of cDNA encoding urinary stone protein, which is identical to osteopontin. Biochem. Biophys. Res. Commun. (1992) 184: 859-864. Kohri, K., et al.
2. Phosphate is a specific signal for induction of osteopontin-gene expression. Proc. Nat. Acad. Sci. (2000) 97: 8352-8357. Beck, G. R., Jr.; Zerler, B.; Moran, E.
3. Osteopontin as a potential diagnostic biomarker for ovarian cancer. JAMA (2002) 287 (13): 1671-1679. Kim J-H., et al.
4. Enhanced local production of osteopontin in rheumatoid joints. J Rheumatol. (2002) 29 (10): 2061-2067. Ohshima S., et al.

ADDITIONAL INFORMATION

• Full Name: Osteopontin N-Half ELISA Kit
• Reactivity: Human
• Sample Type: Serum, Tissue Homogenates, Biological Fluids, Plasma
• Sensitivity:  0.094 ng/ml

OTHER RELATED ELISA KITS

• Human Osteopontin ELISA Kit
• Mouse Osteopontin ELISA Kit
• Intact PTH ELISA Kit

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