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; Judy Aronson; Robert E. Shope; James F. Leary; David G. Gorenstein

doi:10.1093/nar/gng054

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, Judy Aronson, Robert E. Shope, James F. Leary, David G. Gorenstein

Pathology

Research output: Contribution to journal › Article › peer-review

71 Scopus citations

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 language	English (US)
Pages (from-to)	e54
Journal	Nucleic acids research
Volume	31
Issue number	10
DOIs	https://doi.org/10.1093/nar/gng054
State	Published - May 15 2003

ASJC Scopus subject areas

Genetics

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title = "Immunofluorescence assay and flow-cytometry selection of bead-bound aptamers.",

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.",

author = "Xianbin Yang and Xin Li and Prow, {Tarl W.} and Reece, {Lisa M.} and Bassett, {Suzanne E.} and Luxon, {Bruce A.} and Herzog, {Norbert K.} and Judy Aronson and Shope, {Robert E.} and Leary, {James F.} and Gorenstein, {David G.}",

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T1 - Immunofluorescence assay and flow-cytometry selection of bead-bound aptamers.

AU - Yang, Xianbin

AU - Li, Xin

AU - Prow, Tarl W.

AU - Reece, Lisa M.

AU - Bassett, Suzanne E.

AU - Luxon, Bruce A.

AU - Herzog, Norbert K.

AU - Aronson, Judy

AU - Shope, Robert E.

AU - Leary, James F.

AU - Gorenstein, David G.

PY - 2003/5/15

Y1 - 2003/5/15

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AB - 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.

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

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