Description
The device is a colorimetric sandwich ELISA. Samples are diluted in assay diluent and 100uL diluted sample is transferred to the microtiter wells and incubated at room temperature for 60 minutes. During this first incubation C4d in the sample is captured by the anti-C4d-Neo monoclonal antibody, pre-coated on the surface of the microtiter wells. After washing to remove unbound material, a second horseradish peroxidase (HRP) labelled monoclonal antibody, that binds to both allelic variants of C4d (A and B), is added the to the well. After incubation for 30 minutes the wells are washed again, and a substrate is added and incubated. The color development is stopped after 30 minutes, and the color is measured in a spectrophotometer. The color is directly proportional to the amount of C4d bound to the well. The amount of C4d is determined by comparison to the color development of the calibrator samples