I have some experience with in situ PLA, which is a technique developed by O. Söderberg, to detect two proteins in proximity. This technique was published in a widely cited paper (Nat Methods. 2006 Dec;3(12):995-1000) in which some notions about triple PLA were exposed. I recently read a paper about the triple PLA coming from Schallmeiner, E. I´d like to ascertain the experience about the utility of this recently developed technique.
Hi Salvador,
There is a research group in Montreal that detect assembly of 2-4 protein partners in a heteromeric recepter complex.
Here is the PMID: 25053617. The chemokine CXC4 and CC2 receptors form homo- and heterooligomers that can engage their signaling G-protein effectors and βarrestin.
It works using a hybrid of BRET (bioluminescence resonance energy transfer) and PCA (protein-fragments complementation assay).
Thank you for your response Vincent. I did not know this technique at all so I will read the paper with interest. However, as I read in the abstract of the paper you mention they do perform the assays in HEK cells, not in tissue. I would like to detect heterotrimers in fixed tissue in order to explore the presence of heterotrimers in certain regions of the brain.
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