Bovine Haptoglobin ELISA Assay

$580.00

The Bovine Haptoglobin ELISA Assay Kit is a highly sensitive two-site enzyme linked immunoassay (ELISA) for measuring Haptoglobin in biological samples of cattle. The Bovine Haptoglobin kit does not cross-react with Sheep Haptoglobin. If testing sheep samples refer to SHP91-K01, Sheep Haptoglobin ELISA kit.

SKU: BCH61-K01 Categories: , , Tags: , ,

Bovine Haptoglobin ELISA Assay

The Bovine Haptoglobin ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 6.258 ng/ml
Dynamic Range: 15.6 ng/ml- 1000 ng/ml
Incubation Time: 40 minutes
Sample Type: Plasma, Serum
Sample Size: 100 μL
Alternative Names: Cow Haptoglobin ELISA


Assay Background

Acute phase proteins are plasma proteins which increase in concentration following infection, inflammation or trauma. The first acute phase protein to be recognized was discovered in humans by Tillet and Frances in 1930. Haptoglobin (Hp) is a heterogeneous plasma protein mostly synthesized by the liver. The haptoglobin monomer consist of two heavy chains, beta chains (40 kD) and two light chains, alpha chains, alpha 1 (9 kD) and alpha 2 (16 kD) that are linked disulfide bonds. The three major haptoglobin types are; Hp1-1 which is monomeric (98kD), Hp1-2 is polymeric at about 200 kD, and Hp2-2 at about 400 kD. The levels in serum rise quickly following acute tissue damage within 24 to 48 hours and also fall very rapidly once the stimulus is removed. In fact, Hp level are decreased in hemolytic anemia. Hp has a high affinity for hemoglobin (Hb) and its function appears to be to prevent loss of Hb in urine which would lead to loss of iron. Investigations over the past few years have shown that quantification of Hp in plasma or serum can provide valuable diagnostic information in the detection, prognosis, and monitoring of disease not only in humans, but in companion animals and farm herds as well.


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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Additional Information

Assay Principle


The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the Haptoglobin present in samples reacts with the anti-Haptoglobin antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-Hp antibodies conjugated with horseradish peroxidase (HRP) are added. These enzyme-labeled antibodies form complexes with the previously bound Hp. 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 Hp in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of Hp in the test sample. The quantity of Hp in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.

Typical Standard Curve


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