Quantitation of Immunoglobulin E Using Fluorescence Assay
Biological, Environmental, and Earth Sciences
The following paper describes an experiment to fluorescently tag immunoglobulin E (IgE) with Alexa Fluor® 488 Dye carboxylic acid, tetrafluorophenyl (TFP) ester and measure known concentrations of the dye-protein complex using a Biotek Synergy 2 Micro-plate reader to determine its fluorescence intensity. IgE is a human antibody that mediates immediate hypersensitivity reactions and aids the immune system in defending the body against parasitic infection. Fluorescence is the emission of light from a substance that has been exposed to electromagnetic radiation. Electrons in this substance emit light when they return to their ground state following excitement via electromagnetic bombardment. The characteristics and functions of IgE and fluorescence are discussed followed by a detailed protocol on the tagging of Alexa Fluor® 488 Dye to IgE. The fluorescence is successfully measured using the micro-plate reader on samples with known concentrations ranging between 20 μg/dL to 100 μg/dL using a Gen5 computer software program. Lastly, the author explores how the experiment could be implemented into the clinical setting along with how the procedure could be expanded using fluorescence polarization.
(2014). Quantitation of Immunoglobulin E Using Fluorescence Assay. The Chemist, 87(2), 1-7.
Available at: https://aquila.usm.edu/fac_pubs/16196