purification
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Purification Carm1-130-487Bold Text
Sur 1l baculo. Sonication dans 50ml 50mM TrisHCl pH8, 250mM NaCl, 5% glycerol, 5mM tcep, 0.01% NP40 + anti-protéases. Centrifugation 30mn 18000rpm. Incubation EB + 2ml billes GSH-sépharose o/n 4°C. Lavages billes 2x15ml tampon. Digest tev 4h 30°C dans 2ml. Récup surnageant v=2ml, lavage 1 v=2ml, lavage 2 v=1ml. Pool S+L et concentration jusqu’à v=1.5ml. Injection sur GF S200 16/60 et élution 1ml/mn 20mM TrisHCl pH7.5, 50mM NaCl, 1mM tcep.
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