RCA-enhanced protein detection arrays.

Imagen de Paul Lizardi
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TítuloRCA-enhanced protein detection arrays.
Publication TypeJournal Article
Year of Publication2006
AutoresHaab, BB, Lizardi, PM
JournalMethods Mol Biol
Volume328
Pagination15-29
Date Published2006
ISSN1064-3745
Palabras claveAnimals, Antibodies, Fluorescent Dyes, Haptens, Humans, Immunoassay, Oligonucleotides, Protein Array Analysis, Proteins, Proteomics
Abstract

There are many instances in which it is desirable to generate profiles of the relative abundance of a multiplicity of protein species. Examples include studies in embryonic development, immunobiology, drug responses, cancer biology, biomarkers, and so on. Microarray formats provide a convenient, high-throughput vehicle for generating such profiles, and the repertoire of proteins that can be measured is growing continuously as larger panels of specific antibodies become available. Here we describe methods for the use of antibody microarrays, whereby the detection of specifically bound antigens is enhanced by rolling circle amplification (RCA). RCA-enhanced protein detection on antibody microarrays provides a means for rapid protein profiling at high sensitivity. The set of RCA reagents remains unchanged for different microarray formats and compositions, and signal readout is performed using standard fluorescent dyes and scanners. The method is sensitive enough for the most challenging applications, such as the detection of low-abundance components of human serum.

DOI10.1385/1-59745-026-X:15
Alternate JournalMethods Mol. Biol.
PubMed ID16785639
Grant ListCA81671 / CA / NCI NIH HHS / United States
P30 DK 34989 / DK / NIDDK NIH HHS / United States
U01 CA084986 / CA / NCI NIH HHS / United States