- Protocol to quantify palmitoylation of cysteines in budding yeast.
Protocol to quantify palmitoylation of cysteines in budding yeast.
STAR protocols (2022-01-04)
Yuqing Lei, Jiangli Zhu, Huihui Li, Eryan Kong, Kefeng Lu
PMID34977681
ABSTRACT
Palmitoylation is a special kind of lipid modification that targets proteins to membranes. This protocol introduces the acyl-biotin exchange (ABE) assay to determine the palmitoylation of protein cysteines in yeast. Palmitoylation is exchanged by biotinylated compounds so that the palmitoyl proteins can be affinity-purified for downstream assay by western blot. This protocol is easy to perform and can be applied to other biological sources with slight modifications. This protocol is limited to the detection of cysteine-based palmitoylation. For complete details on the use and execution of this profile, please refer to Lei et al. (2021).
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Anti-19S regulator non-ATPase subunit S5a/Rpn10 Antibody, clone S5a-18, clone S5a-18, Upstate®, from mouse