Sigma-Aldrich Smc# Human Ige Immunoassay Kit

The SMC Human IgE 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 immunoassay system detects analytes using Single Molecule Counting (SMC) technology.

The Single Molecule Counting (TM=["SMC"]) Human IgE High Sensitivity Immunoassay uses a quantitative fluorescent sandwich immunoassay technique to measure IgE in human serum and plasma samples. The user coats a capture antibody specific for human IgE onto a 96-well microplate. After blocking the plate, the user pipettes standards and samples into the coated microplate wells. During incubation, the IgE 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 IgE that has been captured onto the plate, thus completing the immunosandwich. 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 TM=["SMCxPRO"] System where the labeled molecules are detected and counted. The number of fluor-labeled detection antibodies counted is directly proportional to the amount of IgE present in the sample when captured. The amount of IgE in unknown samples is interpolated from a standard curve.

This assay does not show cross-reactivity against immunoglobulin subclasses IgA, IgD, IdE, IgG1, IgG2, IgG3, or IgG4 from in-house testing

Minimum of 5 samples spiked and diluted (at least 3 dilutions) with acceptable recovery between 86-89%

Assay Plate Coating Buffer Blocking Buffer Standard Diluent IgM Detection Antibody IgM Standard SMC^® Human IgM Immunoassay Kit Instructions 10X Wash Buffer Buffer C Elution Buffer B Buffer 3 IgM Coating Antibody

SMC is a registered trademark of Singulex

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.