Description
Principle of the Assay: The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the Albumin present in samples reacts with the anti-Albumin antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-Alb antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound ALB. 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 Alb in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of Alb in the test sample. The quantity of Alb in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.
Background: Albumin (Alb) is an amazing polyfunctional protein contributing to homeostasis through mechanisms of hemodynamics, transport and nutrition. Albumin is found both intra and extravascularly in all mammals and many lower vertebrates. It is a molecule of about 67,000 daltons, synthesized by the liver. Normally only very trace amounts of albumin escape reabsorption by kidney glomeruli and is excreted into the urine. Many occult diseases can cause kidney damage which may result in excessive amounts of serum proteins, including albumin, to be excreted by the kidney and into the urine. This ELISA kit can be used to measure albumin in serum, tissue extracts and other biological fluids