Enzyme Immunoassay for the Quantitative Determination of Beta-2 Microglobulin (B2MG) Concentration in Human Serum, Plasma and Urine

Product Description
Human â-2 Microglobulin (B2MG) is an 11.8 kD protein identical to the light chain of the HLA-A, - B, and - C antigen. B2MG is expressed on nucleated cells, and is found at low levels in the serum and urine of normal individuals. B2MG concentrations are increased in inflammatory diseases, some viral diseases, renal dysfunction, and autoimmune diseases. A number of publications are available which explain the interpretation of B2MG serum levels in assessing the status of individuals with various clinical conditions.

Principle
The B2MG ELISA test is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a unique monoclonal antibody directed against a distinct antigenic determinant on the intact â-2 Microglobulin molecule. Mouse monoclonal anti- B2MG antibody is used for solid phase immobilization (on the microtiter wells).
A sheep anti-B2MG antibody is in the antibody-enzyme (horseradish peroxidase) conjugate solution. The diluted test sample is allowed to react first with the immobilized antibody for 30 minutes at 37°C. The sheep anti-B2MG-HRP conjugate is then added and reacted with the immobilized antigen for 30 minutes at 37°C, resulting in the B2MG molecules being sandwiched between the solid phase and enzymelinked antibodies.
The wells are washed with water to remove unbound-labeled antibodies. A solution of TMB Reagent is added and incubated for 20 minutes at room temperature, resulting in the development of a blue color. The color development is stopped with the addition of Stop Solution, changing the color to yellow.
The concentration of B2MG is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450nm.

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