Using linkage-specific monoclonal antibodies to analyze cellular ubiquitylation

Kim Newton; Marissa L Matsumoto; Ronald E Ferrando; Katherine E Wickliffe; Michael Rape; Robert F Kelley; Vishva M Dixit

doi:10.1007/978-1-61779-474-2_13

Using linkage-specific monoclonal antibodies to analyze cellular ubiquitylation

Methods Mol Biol. 2012:832:185-96. doi: 10.1007/978-1-61779-474-2_13.

Authors

Kim Newton¹, Marissa L Matsumoto, Ronald E Ferrando, Katherine E Wickliffe, Michael Rape, Robert F Kelley, Vishva M Dixit

Affiliation

¹ Physiological Chemistry Department, Genentech, Inc., South San Francisco, CA, USA.

PMID: 22350886
DOI: 10.1007/978-1-61779-474-2_13

Abstract

Antibodies that specifically recognize polyubiquitin chains containing ubiquitins linked at a particular lysine residue are powerful tools for interrogating endogenous protein modifications. Here, we describe protocols for revealing K11-, K48-, and K63-linked polyubiquitin chains by western blotting, immunoprecipitation, or immunostaining.

MeSH terms

Antibodies, Monoclonal / immunology*
Blotting, Western / methods
Immunoprecipitation / methods
Polyubiquitin / chemistry*
Polyubiquitin / immunology*
Polyubiquitin / metabolism
Protein Processing, Post-Translational
Ubiquitin-Activating Enzymes
Ubiquitin-Conjugating Enzymes
Ubiquitin-Protein Ligases
Ubiquitination

Substances

Antibodies, Monoclonal
Polyubiquitin
Ubiquitin-Conjugating Enzymes
Ubiquitin-Protein Ligases
Ubiquitin-Activating Enzymes