TY - JOUR
T1 - Proximity Ligation Assay for Detecting Protein-Protein Interactions and Protein Modifications in Cells and Tissues in Situ
AU - Hegazy, Marihan
AU - Cohen-Barak, Eran
AU - Koetsier, Jennifer L.
AU - Najor, Nicole A.
AU - Arvanitis, Constadina
AU - Sprecher, Eli
AU - Green, Kathleen J.
AU - Godsel, Lisa M.
N1 - Publisher Copyright:
© 2020 Wiley Periodicals LLC.
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Biochemical methods can reveal stable protein-protein interactions occurring within cells, but the ability to observe transient events and to visualize the subcellular localization of protein-protein interactions in cells and tissues in situ provides important additional information. The Proximity Ligation Assay® (PLA) offers the opportunity to visualize the subcellular location of such interactions at endogenous protein levels, provided that the probes that recognize the target proteins are within 40 nm. This sensitive technique not only elucidates protein-protein interactions, but also can reveal post-translational protein modifications. The technique is useful even in cases where material is limited, such as when paraffin-embedded clinical specimens are the only available material, as well as after experimental intervention in 2D and 3D model systems. Here we describe the basic protocol for using the commercially available Proximity Ligation Assay™ materials (Sigma-Aldrich, St. Louis, MO), and incorporate details to aid the researcher in successfully performing the experiments.
AB - Biochemical methods can reveal stable protein-protein interactions occurring within cells, but the ability to observe transient events and to visualize the subcellular localization of protein-protein interactions in cells and tissues in situ provides important additional information. The Proximity Ligation Assay® (PLA) offers the opportunity to visualize the subcellular location of such interactions at endogenous protein levels, provided that the probes that recognize the target proteins are within 40 nm. This sensitive technique not only elucidates protein-protein interactions, but also can reveal post-translational protein modifications. The technique is useful even in cases where material is limited, such as when paraffin-embedded clinical specimens are the only available material, as well as after experimental intervention in 2D and 3D model systems. Here we describe the basic protocol for using the commercially available Proximity Ligation Assay™ materials (Sigma-Aldrich, St. Louis, MO), and incorporate details to aid the researcher in successfully performing the experiments.
KW - Duolink™
KW - in situ tissue staining
KW - post-translational modification
KW - protein-protein interaction
KW - proximity ligation assay®
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U2 - 10.1002/cpcb.115
DO - 10.1002/cpcb.115
M3 - Article
C2 - 33044803
AN - SCOPUS:85092944389
SN - 1934-2500
VL - 89
SP - e115
JO - Current Protocols in Cell Biology
JF - Current Protocols in Cell Biology
IS - 1
ER -