Characterizing proteins and their interactions in cells and tissues using the in situ proximity ligation assay

O Söderberg, KJ Leuchowius, M Gullberg, M Jarvius… - Methods, 2008 - Elsevier
O Söderberg, KJ Leuchowius, M Gullberg, M Jarvius, I Weibrecht, LG Larsson, U Landegren
Methods, 2008Elsevier
The activity of proteins is typically regulated by secondary modifications and by interactions
with other partners, resulting in the formation of protein complexes whose functions depend
on the participating proteins. Accordingly, it is of central importance to monitor the presence
of interaction complexes as well as their localization, thus providing information about the
types of cells where the proteins are located and in what sub-cellular compartment these
interactions occur. Several methods for visualizing protein interactions in situ have been …
The activity of proteins is typically regulated by secondary modifications and by interactions with other partners, resulting in the formation of protein complexes whose functions depend on the participating proteins. Accordingly, it is of central importance to monitor the presence of interaction complexes as well as their localization, thus providing information about the types of cells where the proteins are located and in what sub-cellular compartment these interactions occur. Several methods for visualizing protein interactions in situ have been developed during the last decade. These methods in most cases involve genetic constructs, and they have been successfully used in assays of living cell maintained in tissue culture, but they cannot easily be implemented in studies of clinical specimens. For such samples, affinity reagents like antibodies can be used to target the interacting proteins. In this review we will describe the in situ proximity ligation assays (in situ PLA), a method that is suitable for visualizing protein interactions in both tissue sections and in vitro cell lines, and we discuss research tasks when this or other method may be selected.
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