title: Using dot blot with immunochemical detection to evaluate global changes in SUMO-2/3 conjugation.
abstract: Small ubiquitin-related modifier-2/3 (SUMO-2/3) is a member of the ubiquitin-like (Ubl) protein family. Conjugation of SUMO-2/3 to target proteins is influenced by various stress conditions and chemical inhibitors. SUMO-2/3 conjugation may serve as a neuroprotective mechanism and may play a role in protein quality control. A method for screening global changes in SUMO-2/3 conjugation would facilitate further research of SUMO-2/3 cellular function. Here we show that dot blot with immunochemical detection allows evaluation of changes in global cellular SUMO-2/3 conjugation and offers an alternative to more laborious Western blot analysis. The method is based on a change of SUMO-2/3 signal intensity upon its conjugation. The dot blot analysis presented here is a time-saving method that enables screening of large numbers of samples and easy statistical evaluation of the results.
authors: Department of Biochemistry and Microbiology, Institute of Chemical Technology, Prague, Czech Republic.
authors: Markéta
authors: M
authors: Častorálová
authors: Department of Biochemistry and Microbiology, Institute of Chemical Technology, Prague, Czech Republic.
authors: Tomáš
authors: T
authors: Ruml
authors: Department of Biochemistry and Microbiology, Institute of Chemical Technology, Prague, Czech Republic.
authors: Zdeněk
authors: Z
authors: Knejzlík
mesh_terms: D015153:Blotting, Western; D057809:HEK293 Cells; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D025841:Small Ubiquitin-Related Modifier Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D014452:Ubiquitins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism
chemical_list: C117909:SUMO2 protein, human; C492189:SUMO3 protein, human; D025841:Small Ubiquitin-Related Modifier Proteins; D014452:Ubiquitins
keywords: SUMO-2/3; SUMO-2/3 conjugation; dot blot