Immunoprecipitation Using Mono-ADP-Ribosylation-Specific Antibodies

Helen Dauben; Ivan Matic

doi:10.1007/978-1-0716-2891-1_9

Immunoprecipitation Using Mono-ADP-Ribosylation-Specific Antibodies

Methods Mol Biol. 2023:2609:135-146. doi: 10.1007/978-1-0716-2891-1_9.

Authors

Helen Dauben¹, Ivan Matic^{2

3}

Affiliations

¹ Research Group of Proteomics and ADP-ribosylation Signaling, Max Planck Institute for Biology of Ageing, Cologne, Germany. hdauben@age.mpg.de.
² Research Group of Proteomics and ADP-ribosylation Signaling, Max Planck Institute for Biology of Ageing, Cologne, Germany. imatic@age.mpg.de.
³ Cologne Excellence Cluster for Stress Responses in Ageing-Associated Diseases (CECAD), University of Cologne, Cologne, Germany. imatic@age.mpg.de.

PMID: 36515834
DOI: 10.1007/978-1-0716-2891-1_9

Abstract

Immunoprecipitation is an essential methodology for enriching and purifying targeted proteins and peptides for in-depth analysis by any number of further techniques, from Western blotting to mass spectrometry (MS). Historically, the posttranslational modification ADP-ribosylation (ADPr) has been studied mainly in its polymerized form (poly-ADPr), but recent studies support the abundance and physiological relevance of mono-ADPr. Here, we describe several approaches to enrich mono-ADP-ribosylated proteins and peptides using mono-ADPr-specific antibodies, which can be tailored to a desired target and mode of downstream analysis.

Keywords: ADP-ribosylation; Antibodies; Cross-linking; Histones; Immunoprecipitation; Protein of interest.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ADP-Ribosylation*
Antibodies / metabolism
Immunoprecipitation
Peptides / chemistry
Protein Processing, Post-Translational*
Proteins / metabolism

Substances

Proteins
Peptides
Antibodies