No answer to the lack of specificity: mouse monoclonal antibody targeting the angiotensin II type 1 receptor AT1 fails to recognize its target

Abstract : Numerous antibodies targeting G protein-coupled receptors (GPCRs) have been described as non-specific among the polyclonalantibodies against angiotensin II type 1 receptor (AT1). We have tested the newly developed AT1 receptor mouse monoclonalantibody for its specificity. Human embryonic kidney (HEK293) cells, which do not endogenously express AT1 receptor, weretransfected in order to overexpress a fluorescently labeled enhanced green fluorescent protein (EGFP)-tagged human AT1receptor. Western blot and immunofluorescence assays were performed to test the specificity of the Santa Cruz monoclonalantibody sc-57036. These results were compared to the ones obtained with the polyclonal sc-1173 anti-AT1 receptor antibodiesthat have already been described as non-specific. While the positive controls using GFP antibodies detected the EGFP-taggedAT1 receptor, both polyclonal and monoclonal anti-AT1 receptor antibodies failed to specifically recognize the correspondingband by Western blot, as similar bands were revealed in either transfected or non-transfected cells. It also failed to detect AT1receptor in immunofluorescence experiments. The lack of target recognition of the monoclonal AT1 receptor antibody in ourexperimental conditions suggests that this antibody could give misleading results such as misidentification of the protein. To ourknowledge, no specific antibodies targeting AT1 receptors have been developed so far and the field is thus in need of newtechnical developments.
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Marie-Lynda Bouressam, Isabelle Lartaud, François Dupuis, Sandra Lecat. No answer to the lack of specificity: mouse monoclonal antibody targeting the angiotensin II type 1 receptor AT1 fails to recognize its target. Naunyn-Schmiedeberg's Archives of Pharmacology, Springer Verlag, 2018, ⟨10.1007/s00210-018-1522-4⟩. ⟨hal-01816221⟩

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