Canine Hemoglobin ELISA Assay
The Canine Hemoglobin ELISA Assay is For Research Use Only
Size: 1×96 wells
Sensitivity: 3.107 ng/ml
Dynamic Range: 12.5 ng/ml – 400 ng/ml
Incubation Time: 100 minutes
Sample Type: Plasma, Serum
Sample Size: 100 μL
The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the Hemoglobin present in samples reacts with the anti-Hemoglobin antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-Hemoglobin antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound Hemoglobin. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromogenic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of Hemoglobin in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of Hemoglobin in the test sample. The quantity of Hemoglobin 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.