Cite as: Cold Spring Harb. Protoc.; 2008; doi:10.1101/pdb.ip62

This Protocol
Right arrow Full Text
Right arrow Update/discuss this protocolDiscussion icon
Right arrow Alert me when this protocol is cited
Right arrow Alert me when comments are published
Right arrow Alert me if a correction is posted
Services
Right arrow Similar protocols in this database
Right arrow Alert me to new releases of protocols
Right arrow Save to Personal Folders
Right arrow Download to citation manager
Right arrow Printer-friendly versionPrinter-friendly version
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Link, A. J.
Right arrow Articles by LaBaer, J.
Right arrow Search for Related Content
PubMed
Right arrow Articles by Link, A. J.
Right arrow Articles by LaBaer, J.
Related Collections
Right arrow Protein: Protein Interactions
Right arrow CSHL Proteomics Course
Right arrow Protein: Protein Interactions, general
Right arrow Protein Identification and Analysis
Right arrow High-Throughput Analysis, general
Right arrow Molecular Biology, general
Right arrow cDNA
Right arrow Tagging Proteins
Right arrow Proteins and Proteomics, general
Right arrow Immunodetection
Right arrow Proteomics
Right arrow Microarrays
Right arrow Microarrays, general
Right arrow Generation of Probes for Microarrays
Right arrow Spotted Glass Microarrays
Right arrow Proteome Analysis
Right arrow High-throughput Analysis of Protein:Protein Interactions
Right arrow Expression Libraries
Right arrowRelated Protocols
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

information_panelInformation Panel

Using the Nucleic Acid Programmable Protein Array (NAPPA) for Identifying Protein-Protein Interactions: General Guidelines

Andrew J. Link1 and Joshua LaBaer2,3

1 Vanderbilt University School of Medicine, Nashville, TN 37232-8575, USA
2 Harvard University School of Medicine, Harvard Institute of Proteomics, Cambridge, MA 02141-2023, USA

3Corresponding author (joshua_labaer{at}hms.harvard.edu)


INTRODUCTION

The Nucleic Acid Programmable Protein Array (NAPPA) approach for producing protein microarrays uses cell-free extracts to transcribe and translate cDNAs encoding target proteins directly onto glass slides. Following array preparation, the identification of protein interactions using NAPPA can be accomplished by either of two general schemas. The first is to probe an expressed NAPPA slide with a purified protein of interest (the query protein) and look for interactors. Signals of these interactions can be detected either by directly labeling the query protein or by using a labeled antibody either to the protein itself or to a tag on the protein. This approach works well when there is access to the purified protein, and it has the advantage that users can test query protein binding with and without post-translational modifications or under a variety of binding conditions. The second schema entails coexpressing the query protein on the NAPPA slide at the same time that all the target proteins are expressed. This article describes the advantages of using a coexpressed query protein and provides advice on choosing a suitable epitope tag.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?

Related Protocols

Construction of Nucleic Acid Programmable Protein Arrays (NAPPA) 1: Coating Glass Slides with Amino Silane
Andrew J. Link and Joshua LaBaer
Cold Spring Harb Protoc 2008: 5056. [Abstract] [Full Text]

Construction of Nucleic Acid Programmable Protein Arrays (NAPPA) 2: Preparing Bacterial Cultures in a 96-Well Format
Andrew J. Link and Joshua LaBaer
Cold Spring Harb Protoc 2008: 5057. [Abstract] [Full Text]

Construction of Nucleic Acid Programmable Protein Arrays (NAPPA) 3: Isolating DNA Plasmids in a 96-Well Plate Format
Andrew J. Link and Joshua LaBaer
Cold Spring Harb Protoc 2008: 5058. [Abstract] [Full Text]

Construction of Nucleic Acid Programmable Protein Arrays (NAPPA) 4: DNA Biotinylation, Precipitation, and Arraying of Samples
Andrew J. Link and Joshua LaBaer
Cold Spring Harb Protoc 2008: 5059. [Abstract] [Full Text]

Construction of Nucleic Acid Programmable Protein Arrays (NAPPA) 5: Expressing Proteins on NAPPA Slides
Andrew J. Link and Joshua LaBaer
Cold Spring Harb Protoc 2008: 5060. [Abstract] [Full Text]

Construction of Nucleic Acid Programmable Protein Arrays (NAPPA) 6: Detecting Proteins on NAPPA Slides
Andrew J. Link and Joshua LaBaer
Cold Spring Harb Protoc 2008: 5061. [Abstract] [Full Text]

Construction of Nucleic Acid Programmable Protein Arrays (NAPPA) 7: Detecting DNA on NAPPA Slides
Andrew J. Link and Joshua LaBaer
Cold Spring Harb Protoc 2008: 5062. [Abstract] [Full Text]

Using the Nucleic Acid Programmable Protein Array (NAPPA) for Identifying Protein-Protein Interactions. Protocol 1: Coexpression of Query Protein on NAPPA Slides
Andrew J. Link and Joshua LaBaer
Cold Spring Harb Protoc 2008: 5108. [Abstract] [Full Text]

Using the Nucleic Acid Programmable Protein Array (NAPPA) for Identifying Protein-Protein Interactions. Protocol 2: Detection of Query Proteins on NAPPA Slides
Andrew J. Link and Joshua LaBaer
Cold Spring Harb Protoc 2008: 5109. [Abstract] [Full Text]



This article has been cited by other articles:


Home page
 Cold Spring Harb ProtocHome page
A. J. Link and J. LaBaer
Using the Nucleic Acid Programmable Protein Array (NAPPA) for Identifying Protein-Protein Interactions. Protocol 1: Coexpression of Query Protein on NAPPA Slides
Cold Spring Harb Protoc, December 1, 2008; 2008(13): pdb.prot5108 - pdb.prot5108.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
A. J. Link and J. LaBaer
Using the Nucleic Acid Programmable Protein Array (NAPPA) for Identifying Protein-Protein Interactions. Protocol 2: Detection of Query Proteins on NAPPA Slides
Cold Spring Harb Protoc, December 1, 2008; 2008(13): pdb.prot5109 - pdb.prot5109.
[Abstract] [Full Text]