Intended Use:The total Human IgG test kits are a highly sensitive two-site enzyme linked immunoassay (ELISA) for measuring IgG in serum or plasma of humans.
Test Principle:The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the IgG present in samples reacts with the anti-IgG antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-IgG antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound IgG. 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 IgG in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of IgG in the test sample. The quantity of IgG in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.
Kit Components: (Quantities sufficient for 1x96 determinations)Diluent Concentrate (Running Buffer)One bottle containing 1x50ml of a 5X concentrated diluent running buffer.
Wash solution concentrateOne bottle containing 1x50l of a 20X concentrated wash solution.
Enzyme-antibody conjugate 100XOne vial containing 1x150ul of affinity purified anti-Human IgG antibody conjugated with horseradish peroxidase in a stabilizing buffer.
Chromogen-substate solution One vial containing 1x12ml of 3,3’,5,5’-tetramethybenzidine (TMB) and hydrogen peroxide in citric acid buffer at pH 3.3.
Stop solution One vial containing 1x12ml 0.3M sulfuric acid.
Anti-human IgG ELISA micro plateTwelve removable eight (8) well microwell strips in well holder frame. Each well is coated with affinity purified anti-Human IgG.
Human IgG calibratorOne vial containing Human IgG calibrator.
Storage and Stability:Store all components at 4°C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
