Reverse-Phase Protein Array: Technology, Application, Data Processing, and Integration.
post-translational
protein array analysis
proteomics
software
validation study
Journal
Journal of biomolecular techniques : JBT
ISSN: 1943-4731
Titre abrégé: J Biomol Tech
Pays: United States
ID NLM: 100888641
Informations de publication
Date de publication:
04 2021
04 2021
Historique:
entrez:
24
5
2021
pubmed:
25
5
2021
medline:
29
10
2021
Statut:
ppublish
Résumé
Reverse-phase protein array (RPPA) is a high-throughput antibody-based targeted proteomics platform that can quantify hundreds of proteins in thousands of samples derived from tissue or cell lysates, serum, plasma, or other body fluids. Protein samples are robotically arrayed as microspots on nitrocellulose-coated glass slides. Each slide is probed with a specific antibody that can detect levels of total protein expression or post-translational modifications, such as phosphorylation as a measure of protein activity. Here we describe workflow protocols and software tools that we have developed and optimized for RPPA in a core facility setting that includes sample preparation, microarray mapping and printing of protein samples, antibody labeling, slide scanning, image analysis, data normalization and quality control, data reporting, statistical analysis, and management of data. Our RPPA platform currently analyzes ∼240 validated antibodies that primarily detect proteins in signaling pathways and cellular processes that are important in cancer biology. This is a robust technology that has proven to be of value for both validation and discovery proteomic research and integration with other omics data sets.
Identifiants
pubmed: 34025221
doi: 10.7171/jbt.21-3202-001
pii: JBT_2021-3202-001
pmc: PMC7861052
doi:
Substances chimiques
Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
15-29Subventions
Organisme : NCI NIH HHS
ID : P30 CA125123
Pays : United States
Organisme : NIEHS NIH HHS
ID : P30 ES030285
Pays : United States
Organisme : NIEHS NIH HHS
ID : P42 ES027725
Pays : United States
Informations de copyright
© Association of Biomolecular Resource Facilities.
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