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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Yang X,Li X,Prow TW,Reece LM,Bassett SE,Luxon BA,Herzog NK,Aronson J,Shope RE,Leary JF,Gorenstein DG

doi

10.1093/nar/gng054

keywords:

subject

Has Abstract

pub_date

2003-05-15 00:00:00

pages

e54

issue

10

eissn

0305-1048

issn

1362-4962

journal_volume

31

pub_type

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