title: Detection and analysis of FAT10 modification.
abstract: FAT10 plays a role in many cellular processes. Nevertheless, only one substrate could be identified so far to which FAT10 becomes covalently attached via a nonreducible isopeptide bond. The identification of additional substrates as well as interaction partners is therefore of great interest. Due to the absence of potent anti-FAT10 antibodies, the detection and identification of interaction partners was hindered so far. We have recently described the generation of a novel monoclonal FAT10 antibody that is suitable for immunoprecipitation of endogenous FAT10 and describe in this chapter the detection of endogenous as well as of His-3×FLAG-tagged FAT10 interacting proteins and conjugates by immunoprecipitation using either this novel anti-FAT10 antibody 4FI or an anti-FLAG affinity gel.
authors.affiliation: Biotechnology Institute Thurgau, Kreuzlingen, Switzerland. annette.aichem@bitg.ch
authors.forename: Annette
authors.initials: A
authors.lastname: Aichem
authors.forename: Marcus
authors.initials: M
authors.lastname: Groettrup
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D002453:Cell Cycle; D059447:Cell Cycle Checkpoints; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D016328:NF-kappa B / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism; D014452:Ubiquitins / Q000032:analysis* / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D016328:NF-kappa B; C109184:UBD protein, human; D025801:Ubiquitin; D014452:Ubiquitins