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Beschreibung

The Single Molecule Counting (SMC(R) Human IgM High Sensitivity Immunoassay uses a quantitative fluorescent sandwich immunoassay technique to measure IgM in human serum and plasma samples. The user coats a capture antibody specific for human IgM onto a 96-well microplate. After blocking the plate, the user pipettes standards and samples into the coated microplate wells. During incubation, the IgM present in the sample binds to the capture antibody on the coated plate. Unbound molecules are washed away during a wash step. The fluor-labeled detection antibody is added to each well and incubated. The detection antibody recognizes and binds to IgM that has been captured onto the plate, thus completing the immunosanwich. Following the final wash step, elution buffer is added and incubated. The elution buffer dissociates the bound protein sandwich from the plate surface, releasing the labeled antibodies. These antibodies are separated during transfer to a final microplate. The reading plate is loaded onto the Erenna or SMCxPRO(R) System where the labeled molecules are detected and counted. The number of fluor-labeled detection antibodies counted is directly proportional to the amount of IgM present in the sample when captured. The amount of IgM in unknown samples is interpolated from a standard curve.The SMC(R) Human IgM High Sensitivity Immunoassay Kit contains all reagents required to perform the assays. This assay takes advantage of a standard immunoassay workflow configured in a standard plate format. Assay protocols are similar to existing sandwich ELISA methods with two key differences: 1) Elution buffer disrupts the sandwich, separating the labeled detection antibody for quantification. 2) The SMC(R)immunoassay system detects analytes using Single Molecule Counting (SMC(R)) technology.