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Low‐SDS Blue native PAGE
Author(s) -
Klodmann Jennifer,
Lewejohann Dagmar,
Braun HansPeter
Publication year - 2011
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201000638
Subject(s) - polyacrylamide gel electrophoresis , gel electrophoresis , protein subunit , biochemistry , chemistry , electrophoresis , arabidopsis , biology , gene , enzyme , mutant
SDS normally is strictly avoided during Blue native (BN) PAGE because it leads to disassembly of protein complexes and unfolding of proteins. Here, we report a modified BN‐PAGE procedure, which is based on low‐SDS treatment of biological samples prior to native gel electrophoresis. Using mitochondrial OXPHOS complexes from Arabidopsis as a model system, low SDS concentrations are shown to partially dissect protein complexes in a very defined and reproducible way. If combined with 2‐D BN/SDS‐PAGE, generated subcomplexes and their subunits can be systematically investigated, allowing insights into the internal architecture of protein complexes. Furthermore, a 3‐D BN/low‐SDS BN/SDS‐PAGE system is introduced to facilitate structural analysis of individual protein complexes without their previous purification.