Using dot blot with immunochemical detection to evaluate global changes in SUMO-2/3 conjugation | Zendy

Markéta Častorálová | Zendy; Tomáš Ruml | Zendy; Zdeněk Knejzlı́k | Zendy

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Using dot blot with immunochemical detection to evaluate global changes in SUMO-2/3 conjugation

Author(s) -

Markéta Častorálová,

Tomáš Ruml,

Zdeněk Knejzlı́k

Publication year - 2012

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/000113925

Subject(s) - western blot , ubiquitin , sumo protein , dot blot , function (biology) , blot , computational biology , chemistry , biochemistry , microbiology and biotechnology , biology , dna , gene

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.

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