SPINE (molecular biology)

Strep protein interaction experiment ( SPINE ) is a biochemical method of detecting protein-protein interactions.

Properties

SPINE which is a variant of the pull-down assays in the detection of a contact of two or more proteins, protein purification by a reference to a strep -tag at a known, most recombinant protein. By cleaning the binding proteins are mitaufgereinigt. Purification is carried out by affinity chromatography (e.g., streptactin ) with a reversible cross-linking of the protein complexes of formaldehyde for fixation of the most temporary bond. By boiling the cross-linked protein complexes in sample buffer networking is canceled. Further separation is then obtained by SDS -PAGE. The identification of binding proteins is accomplished by a subsequent in-gel digestion and analysis by mass spectrometry. The method is adapted to reduce non-specific interactions, due to the hydrophobic effect of proteins.