Purification of Protein-complexes from the Cyanobacterium Synechocystis sp. PCC 6803 Using FLAG-affinity Chromatography
Minna M. Koskela, Petra Skotnicová, Éva Kiss, Roman Sobotka
Abstract
. Soluble proteins or solubilized thylakoids are subjected to a single affinity purification step that utilizes the highly specific binding of FLAG-affinity resin. After an intensive wash, the captured proteins are released from the resin under native conditions using an excess of synthetic 3x FLAG peptide. The protocol allows fast isolation of low abundant protein complexes with a superb purity.
Topics & Concepts
Affinity chromatographySynechocystisProtein purificationFLAG-tagBiochemistryBiogenesisChromatographyChemistryThylakoidMembrane proteinCell fractionationUltracentrifugeMembraneChloroplastEnzymeRecombinant DNAFusion proteinMutantGenePhotosynthetic Processes and MechanismsAlgal biology and biofuel productionProtist diversity and phylogeny