RPPA does not separate proteins according to their molecular weight; the obtained signal is concentrated in one spot. Therefore, antibodies for RPPA need to be highly specific. Antibody testing is an important part of our activity. We have tested so far >2100 commercial antibodies on a panel of cell lines, grown in different conditions in order to activate a maximum of cell signaling pathways. We use a panel of human cell lines and one mouse cell line (NIH-3T3). We can therefore tell whether an antibody recognizes the murine protein, at least in NIH-3T3 cells. This can be useful for projects concerning xenografts or mouse tissue. FWe aim for one major band at the expected molecular weight in Western Blot (or several bands in case of multiple isoforms).
So far we have validated >640 primary antibodies for RPPA.
The list of proteins that we can currently analyze by RPPA can be downloaded here: ValidAntibodies_2018-04-26
For scientists working at Institut Curie: you can connect to our database and visualize our Western Blot results. To do so, connect as a guest to http://parys.curie.fr/.