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 67kD, 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.

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

A1274-92B1: Microtiter Plate, 1x 96 wells (12 removable 8-microwell strips in well holder frame. Each well is coated with purified anti-rat albumin. A1274-92B2: Antibody (HRP), 1x150ul. Undiluted affinity-purified anti-rat albumin antibody conjugated with horseradish peroxidase in a stabilizing buffer *A1274-92B3: Calibrator, 1x 1vial. Lyophilized rat albumin calibrator. A1274-92B4: TMB Substrate, 1x12ml. 3,3’,5,5’- tetramethybenzidine (TMB) and hydrogen peroxide in citric acid buffer at pH 3.3. A1274-92B5: Diluent Concentrate, 5X, 1x50ml. A1274-92B6: Wash Solution Concentrate, 20X, 1x50ml. A1274-92B7: Stop Solution, 1x12ml. 0.3M sulfuric acid. Warning: Avoid contact with skin.

Store *A1274-92B3: Calibrator powder and aliquoted liquid at -20°C. Store other components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.