- Split-miniSOG for Spatially Detecting Intracellular Protein-Protein Interactions by Correlated Light and Electron Microscopy.
Split-miniSOG for Spatially Detecting Intracellular Protein-Protein Interactions by Correlated Light and Electron Microscopy.
Cell chemical biology (2019-08-06)
Daniela Boassa, Sakina P Lemieux, Varda Lev-Ram, Junru Hu, Qing Xiong, Sebastien Phan, Mason Mackey, Ranjan Ramachandra, Ryan Emily Peace, Stephen R Adams, Mark H Ellisman, John T Ngo
PMID31378710
ABSTRACT
A protein-fragment complementation assay (PCA) for detecting and localizing intracellular protein-protein interactions (PPIs) was built by bisection of miniSOG, a fluorescent flavoprotein derived from the light, oxygen, voltage (LOV)-2 domain of Arabidopsis phototropin. When brought together by interacting proteins, the fragments reconstitute a functional reporter that permits tagged protein complexes to be visualized by fluorescence light microscopy (LM), and then by standard as well as "multicolor" electron microscopy (EM) via the photooxidation of 3-3'-diaminobenzidine and its derivatives.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Rp-Adenosine 3′,5′-cyclic monophosphorothioate triethylammonium salt, powder, ≥98% (HPLC)
Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
Sigma-Aldrich
Anti-HA antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution