Test Principle:The Human Albumin ELISA kit is based on the binding of human albumin in samples to two antibodies, one immobilized on the microwells, and the other conjugated to horseradish peroxidase (HRP) enzyme. After a washing step, chromogenic substrate (TMB) is added and color is developed by the enzymatic reaction of HRP on the substrate, which is directly proportional to the amount of Albumin present in the sample. Stopping Solution is added to terminate the reaction, and absorbance at 450nm is then measured using an ELISA microwell reader. The concentration of albumin in samples and control is calculated from a curve of standards containing known concentrations of albumin.
Intended Use:The Human Albumin ELISA Kit is an immunoassay suitable for quantifying circulating serum albumin in humans. The assay can be adapted to measure albumin in other biological fluids and solutions such as plasma, urine, and culture medium with proper control for assay compatibility.
Sensitivity:The antibodies used in this kit have been shown by immunoelectrophoresis and ELISA to react specifically with albumin, and have essentially no reactivity with immunoglobulins or any other human serum proteins.
Serum from Rhesus monkey showed moderate reactivity in the ELISA, but monkey albumin cannot be accurately quantified in this assay. Sera from the following species showed no significant reactivity at 1:100K dilution: mouse, rat, hamster, guinea pig, bovine, porcine, sheep, rabbit, goat and chicken. Fetal bovine serum at 10% also showed no reactivity.
Range:Assay values of albumin in sera from 20 adult humans ranged from 14.1-52.4mg/ml (median: 24.3mg/ml). Each laboratory should determine expected values of its own testing population.
Kit Components:A1274-85A1: Microtiter Plate: 1x96 wellsA1274-85A2: Control (24-46ng/ml), 1x650ulA1274-85A3: Standard (5ng/ml), 1x650ulA1274-85A4: Standard (10ng/ml), 1x650ulA1274-85A5: Standard (20ng/ml), 1x650ulA1274-85A6: Standard (50ng/ml), 1x650ulA1274-85A7: Standard (100ng/ml), 1x650ulA1274-85A8: HRP Conjugate, 100X, 1x150ulA1274-85A9: Sample Diluent, 20X, 1x10mlA1274-85A10: Wash Solution, 100X, 1x10mlA1274-85A11: TMB Substrate, 1x12mlA1274-85A12: Stop Solution (1% H2SO4), 1x12ml
Storage and Stability:Store all components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
