Multiplexing HTRF cytokine assays
We have developed several homogeneous cytokine assays based on time-resolved fluorescence resonance energy transfer (TR-FRET) using Europium cryptate and cross-linked allophycocyanine conjugated antibodies. Taking advantage of homogeneous assays, we have co-incubated the cells with the antibody pair during the stimulation step. The results show that the presence of cells in wells at the time of reading does not alter the results. Furthermore, this experimental procedure avoids cumbersome transfer and storage steps streamlining sample analysis. Finally, it facilitates the follow-up of cytokine production.
Assay Drug Dev Technol. 2003;1(1 Pt 2):181-5.