Anti-IgE Receptor (FcɛR1α) Autoantibody ELISA Kit

• Only 10μL of human serum or plasma is required for anti-human IgE receptor antibody measurement in duplicate.
•  In the case of serum, whole blood should be collected and must be allowed to clot for a minimum of 30 minutes at room temperature before the serum is separated by centrifugation (850 – 1500 x g for 10 minutes). The serum should be separated from the clot within three hours of blood collection and transferred to a clean test tube.
• Serum or plasma samples should be stored at 2 – 8°C if the assay is to be performed within 72 hours.
• Avoid repeated (more than three times) freezing and thawing of the specimen.

This ELISA is designed, developed and produced for the quantitative measurement of human anti-hIgE receptor (FcεR1α) autoantibodies in serum and plasma samples.

Assay calibrators, controls and diluted patient samples are directly added to wells of a microplate that is coated with recombinant human IgE receptor protein. After the first incubation period, anti-IgE receptor antibodies bind to the human IgE receptor protein on the wall of microtiter well and unbound proteins in each microtiter well are washed away. Highly purified Protein A labeled with horseradish peroxidase is then added to each microtiter well. After the second incubation period, a complex of coated human IgE receptor ̶ Anti- hIgE receptor antibody ̶ peroxidase-labeled Protein A is formed. The unbound protein is removed in the subsequent washing step. The wells are then incubated with a substrate solution in a timed reaction and subsequently measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the IgE receptor protein on the wall of the microtiter well is directly proportional to the amount of anti-IgE receptor antibody in the sample. A calibration curve is generated by plotting the absorbance versus the respective anti-IgE receptor concentration for each calibrator on a cubic spline curve fit. The concentration of IgE receptor antibody in test samples is determined directly from this calibration curve.