Quantification of Immune Mediator Levels in Human Mucosal Lining Fluid
Transcrição
Thaw the filter papers containing the absorbed nasal mucosal lining fluid on ice to prevent sample degradation.
The fluid comprises immune mediators, including chemokines and cytokines.
Immerse the filter papers in a buffer. Transfer the contents into tube filters. Centrifuge to elute the mucosal lining fluid from the filter paper.
Add the filtered nasal extract to the assay plate patterned with an array of mediator-specific capture antibodies.
The immune mediators bind to their corresponding antibodies.
Introduce detection antibodies labeled with a ruthenium complex, which bind to their respective mediators.
Add tris-based buffer containing tripropylamine, or TPA.
When a potential is applied across the electrodes, the ruthenium complex and TPA undergo oxidation.
The resulting TPA radical excites the ruthenium complex.
Upon returning to its ground state, the ruthenium complex emits photons.
The measured light intensity is proportional to the specific immune mediator concentration in the mucosal lining fluid.
