Bovine Haptoglobin ELISA Kit

Bovine HatoglobinSample Type: Serum / Plasma
Intended Use: The HAPTOGLOBIN test kits are a highly sensitive two-site enzyme linked immunoassay (ELISA) for measuring HAPTOGLOBIN in serum or plasma of cattle. Introduction: 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 19301. 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 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 well2. Principle of the test: The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the HAPTOGLOBIN present in serum sample reacts with the anti-HAPTOGLOBIN antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound serum proteins by washing anti-Hp antibodies conjugated with horseradish peroxidase (HRP) are added. These enzyme-labeled antibodies form complexes with the previously bound serum 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 serum dilutionStorage and Stability: The expiration date for the package is stated on the box label. 1. DILUENTThe 5X Diluent Concentrate is stable until the expiration date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions should be stored at 4-8C. 2. WASH SOLUTIONThe 20X Wash Solution Concentrate is stable until the expiration date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions can be stored at room temperature (16-25C) or at 4-8C. 3. ENZYME-ANTIBODY CONJUGATEUndiluted horseradish peroxidase anti-Hp conjugate should be stored at 4-8C and diluted immediately prior to use. The working conjugate solution is stable for one day. 4. CHROMOGEN-SUBSTRATE SOLUTIONThe Substrate Solution should be stored at 4-8C and is stable until the expiration date. 5. STOP SOLUTIONThe Stop Solution should be stored at 4-8C and is stable until the expiration date. 6. ANTI-BOVINE HAPTOGLOBIN ELISA MICRO PLATEAnti-Bovine Hp coated wells are stable until the expiration date and should be stored at 4-8C in sealed foil pouch with desiccant pack. 7. BOVINE HAPTOGLOBIN STANDARDSThe lyophilized Bovine Haptoglobin calibrator should be stored at 4C or frozen until reconstituted. The reconstituted calibrator should be aliquoted out and stored frozen (Avoid multiple freeze-thaw cycles). The working standard solutions should be prepared immediately prior to use and are stable for 1 day. Limitation of the Test: 1. Reliable and reproducible results will be obtained when the assay procedure is carried out with a complete understanding of the information contained in the package insert instructions and with adherence to good laboratory practice. 2. Factors that might affect the performance of the assay include proper instrument function cleanliness of glassware quality of distilled or deionized water and accuracy of reagent and sample pipettings.

Haptoglobin combines with free plasma hemoglobin preventing loss of iron through the kidneys and protecting the kidneys from damage by hemoglobin while making the hemoglobin accessible to degradative enzymes.

Secreted.

In the human populations there are two major allelic forms alpha-1 with 83 residues and alpha-2 with 142 residues. These alleles determine the 3 major phenotypes HP*1F/HP*1S and HP*2FS. The two main alleles of HP*1 are called HP*1F (fast) and HP*1S (slow).

Contains 1 peptidase S1 domain [view classification].