Determination of Interaction between NFκB p50 and β-IFN-κB Binding Oligo Using AlphaLISA in HTP Fashion

DOI: 10.4236/jasmi.2013.33022   PDF   HTML     2,245 Downloads   3,945 Views   Citations


NF-κB plays a crucial role in regulating various biological processes including innate and adaptive immunity, inflammation, stress responses, B-cell development, and lymphoid organogenesis. Currently, several assays like electrophoretic mobility shift assay (EMSA), enzyme-linked immunosorbent assay (ELISA), fluorescence resonance energy transfer (FRET) and time-resolved fluorescence resonance energy transfer (TR-FRET) are widely used for studying the NFκB intraction with β-IFN-κB binding oligo. Each of these techniques has varying utility with distinct strengths and weaknesses. We describe a method AlphaLISA to identify NFκB p50 protein and β-IFN-κB binding oligo sequence and interaction is efficient at a given concentration (10 nM) in the EMSA and Biacore’s SPR assays. The method has many advantages such as use of small volume, high throughput (HTP), convenience of sample preparation and data analysis.

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M. Neerathilingam, S. Gandham, F. Patel and M. Nasiruddin, "Determination of Interaction between NFκB p50 and β-IFN-κB Binding Oligo Using AlphaLISA in HTP Fashion," Journal of Analytical Sciences, Methods and Instrumentation, Vol. 3 No. 3, 2013, pp. 173-178. doi: 10.4236/jasmi.2013.33022.

Conflicts of Interest

The authors declare no conflicts of interest.


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