Canine Osteopontin ELISA Assay
The Canine Osteopontin ELISA Assay is For Research Use Only
Size: 1×96 wells
Sensitivity: 0.07 ng/ml
Dynamic Range: 10 ng/ml-0.3125 ng/ml
Incubation Time: 170 minutes
Sample Type: Plasma, Urine
Sample Size: 100 μL
Alternative Names: OPN
Assay Principle
In this assay the Osteopontin present in samples reacts with the anti-Osteopontin antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-OPN antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound OPN. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromo-genic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the OPN concentration in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of OPN. The quantity of OPN in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.
SPECIMEN COLLECTION AND HANDLING
Blood should be collected by venipuncture. The serum should be separated from the cells after clot formation by centrifugation. For plasma samples, blood should be collected into a container with an anticoagulant and then centrifuged. Care should be taken to minimize hemolysis, excessive hemolysis can impact your results. Assay immediately or aliquot and store samples at -20°C. Avoid repeated freeze-thaw cycles.
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