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That rules out, straightforwardly, radioligand or fluorescent binding assays, competition binding against a known ligand, recombinant receptor-expressing lines, functional readouts tied to a defined coupling pathway, and antagonist controls to demonstrate that an effect is receptor-mediated
Tandem mass spectrometry analysis of affinity-purified samples Three micrograms of each fraction or sample were auto-sampler loaded with an UltiMate 3000 HPLC pump onto a vented Acclaim Pepmap 100, 75 m 2 cm, nanoViper trap column (Thermo Fisher Scientific, Cat# 164535) coupled to a nanoViper analytical column (Thermo Fisher Scientific, Cat# 164570, 3 m, 100 , C18, 0.075 mm, 500 mm) with stainless steel emitter tip assembled on the Nanospray Flex Ion Source with a spray voltage of 2000 V
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