Rat Haptoglobin ELISA Assay Kit

$460.00

The Eagle Biosciences Rat Haptoglobin ELISA Assay Kit is a highly sensitive two-site enzyme linked immunoassay (ELISA) for measuring Haptoglobin in biological samples of rats.

SKU: RHP21-K01 Categories: , , , Tags: ,

Rat Haptoglobin ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 1.047 ng/ml
Dynamic Range: 1.95 ng/ml – 125 ng/ml
Incubation Time: 40 minutes
Sample Type: Plasma, Serum, Urine
Sample Size: 100 μL

Product manufactured in the U.S.A

Additional Information

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 in 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 by 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 haptoglobin level in serum rise quickly following acute tissue damage within 24 to 48 hours and also falls 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.

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


Manual

Product Manual