Principle of Assay
This B2MG enzyme linked immunosorbent assay (ELISA) kit uses a technique called quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific for B2MG. Standards or samples are added to the microtiter plate wells and B2MG, if present, will bind to the antibody pre-coated on the wells. A preparation of horseradish peroxidase (HRP)-conjugated B2MG antibody is added to each well. The conjugated antibody will bind to the B2MG immobilized on the plate after incubation. All unbound components will be removed by subsequent washing procedure. Next, 3, 3', 5, 5' tetramethyl-benzidine(TMB), a substrate for HRP is added to each well. The colorimetric reaction is proportional to the B2MG content in each well. The reaction is terminated by the addition of a sulphuric acid solution and the colour change is measured spectrophotometrically at the wavelength of 450nm ± 2nm.
In order to quantify the amount of B2MG present in the sample, this Human B2MG ELISA Kit includes a set of calibration standards (6 standards). The calibration standards should be assayed at the same time as the samples to allow the operator to produce a standard curve of Optical Density (O.D.) versus B2MG concentration (mg/mL). The concentration of B2MG in the samples is then determined by comparing the O.D. of the samples to the standard curve.
