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M. Méthanol and 2. , La membrane est ensuite réhydratée par une solution de PBS-Tween (Phosphate de sodium 10 mM, NaCl 140 mM, Tween-20 0 La membrane est ensuite incubée pendant 1 heure en présence d'anticorps primaires anti-CnrXs, La migration s'effectue à 100 V et 350 mA pendant 35 minutes dans le tampon de transfert saturée 30 minutes dans du PBS-Tween-Lait 5%, p.30000

P. Dans-du and 5. , Après lavage dans du PBS-Tween, la membrane est incubée pendant 1 heure en présence d'anticorps secondaires couplés à une peroxydase Après plusieurs lavages au PBS-Tween, les anticorps s'étant couplés aux protéines d'intérêt sont détectés grâce au réactif ECL (Luminata Forte, Millipore) Le signal de chimioluminescence est détecté par exposition d'un film radiographique

0. Tfa, tampon A) à 100 ?L/min sur une colonne Trap (Michrom Bioresources) Les protéines sont ensuite éluées avec 50% de tampon B (90% d'acétonitrile et 0,0% de TFA) à 300 µL/min sur une colonne HPLC C4. Pour éviter le rééchange, les tampons, les vannes et les colonnes sont conservées à 4°C

. Outre-les-logiciels-de-bruker and . Agilent, Sierra Analytics) a été utilisé pour le traitement des données. 3. Identification des peptides de CnrXs?Nter impliqués dans l'interaction L'expérience HDX-MS est similaire à celle effectuée avec les protéines entières à quelques différences près : Après avoir arrété l'échange H/D avec la solution de quenching, les protéines sont digérées sur une colonne de pepsine immobilisée. Le dessalage s'effectue à l'aide d'une colonne Microtrap (Michrom Bioresources)

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