Osteopontin ELISA Kit (OPN)

INTENDED USE

Human osteopontin ELISA kit is a reliable quantitative method for measuring osteopontin (OPN) in human cell culture media, plasma and urine samples. This assay has a sensitivity detection limit of 3.33 ng/ml.

BACKGROUND

Osteopontin (OPN) contains an Arg-Gly-Asp (RGD) amino acid sequence, this motif is present in; vitronectin, fibronectin and many extra cellular proteins which have the ability to bind members of the integrin family of cell surface receptors. It can be also present in a number of different molecular forms in vivo that can be due to; phosphorylation, glycosylation, susceptibility to proteases, sulfation and differential RNA splicing.

Osteopontin is susceptible to proteolytic fragmentation and there is suggestions that this process may have physiologic importance, for example, a study demonstrated that thrombin treatment can enhance OPN cell adhesive activity, implying that cleavage by thrombin can lead to exposing a cryptic adhesive sequence.

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.

• Pre-Coated Microplate: Wells bound with OPN antigen (O-17) rabbit IgG affinity purify.
• Labelled Antibody (30x Concentrated): Consisting of HRP conjugated to anti-OPN (10A16) mouse IgG.
• Standards: Concentrations 0, 5, 10, 20, 40, 80, 160, 320.0 ng/ml.
• Antibody Labelling Solution.
• TMB Chromogen Solution.
• Stop Solution.
• Wash buffer (40x Concentrated).

SENSITIVITY

The minimum sensitivity detection limit of osteopontin (OPN) using current osteopontin ELISA kit was approximately 3.33 ng/ml. The dynamic range for this assay is 5.0 – 320.0 ng/ml.

ASSAY CHARACTERISTICS

– Recovery (Accuracy): 95.6 – 108.7%
– Intra Assay: 4.4 – 10.1%
– Inter Assay: 7.8 – 9.3%
– Specificity: Human-OPN (100%), Mouse-OPN (0.2%), Rat-OPN (<0.1%).
– Sensitivity: 3.33 ng/ml

REFERENCES

1. Phosphate is a specific signal for induction of osteopontin gene expression. Proc. Nat. Acad. Sci. (200) 97: 8352-8357. Beck, G. R.; Jr. Zerler, B. and Moran, E.
2. 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.
3. Human bone sialoprotein: deduced protein sequence and chromosomal localization. J. Biol. Chem. (1990) 265: 2347-2351. Fisher, L. W.; McBride, O. W.; Termine, J. D. and Young, M. F.

ADDITIONAL INFORMATION

• Full Name: Osteopontin ELISA Kit (OPN)
• Reactivity: Human
• Sample Type: Plasma (EDTA), Cell Culture Media, Urine
• Sensitivity: 3.33 ng/ml

OTHER RELATED ELISA KITS

• 25-OH Vitamin D ELISA Kit
• Osteopontin N-Half ELISA Kit
• Calcitonin ELISA Kit