- sELISA
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Sandwich ELISA - Horse Haptoglobin ELISA Kit (AB190528)
Representative standard curve using ab190528 Haptoglobin Horse ELISA Kit.
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Product details
Abcam's Haptoglobin Horse ELISA kit (ab190528) is an in vitro enzyme-linked immunosorbent assay (ELISA) for the quantitative measurement of Haptoglobin in horse serum and plasma.
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-Haptoglobin antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound Haptoglobin. 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 Haptoglobin in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of Haptoglobin in the test sample. The quantity of Haptoglobin in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.
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.
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Biological function summary
Haptoglobin functions as an antioxidant and an acute phase protein playing roles in inflammation and immune response. It is an essential component of the antioxidant defense system reducing oxidative stress by binding free hemoglobin. Additionally haptoglobin can participate in complex formation with hemoglobin to facilitate the clearance of excess free hemoglobin from circulation. Its expression increases during inflammation under the regulation of cytokines like IL-6.
Pathways
The haptoglobin protein is part of the hemoglobin clearance pathway integral for iron metabolism and homeostasis. This pathway ensures the safe removal of hemoglobin from the blood helping to prevent kidney damage. Haptoglobin is related to other proteins such as CD163 and the liver-derived hepatocellular proteins involved in the clearance process. Also haptoglobin is linked to antioxidant pathways collaborating with oxidative stress regulators to manage reactive oxygen species levels.
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