Immunofluorescence assay and flow-cytometry selection of bead-bound aptamers.

Xianbin Yang, Xin Li, Tarl W. Prow, Lisa M. Reece, Suzanne E. Bassett, Bruce A. Luxon, Norbert K. Herzog, Judith Aronson, Robert E. Shope, James F. Leary, David G. Gorenstein

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Abstract

An immunofluorescence assay was developed to identify proteins specifically binding to oligonucleoside phosphorodithioate (ODN) aptamers from a bead-bound ODN library. Accordingly, NF-kappaB p50 protein was incubated with either bead-bound NF-kappaB consensus sequence or a bead-bound ODN combinatorial library and adsorption was then assessed using a specific primary antibody and a secondary antibody conjugated with Alexa 488 fluorescent dye. This assay avoids any problems related to fluorescently labeling target proteins. The method is straightforward and readily applicable to other transcription factors and proteins, and the feasibility of its application for high-throughput screening of large aptamer bead-based libraries by flow cytometry is demonstrated.

Original languageEnglish (US)
JournalNucleic Acids Research
Volume31
Issue number10
StatePublished - May 15 2003

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ASJC Scopus subject areas

  • Genetics

Cite this

Yang, X., Li, X., Prow, T. W., Reece, L. M., Bassett, S. E., Luxon, B. A., Herzog, N. K., Aronson, J., Shope, R. E., Leary, J. F., & Gorenstein, D. G. (2003). Immunofluorescence assay and flow-cytometry selection of bead-bound aptamers. Nucleic Acids Research, 31(10).