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▪ Slow injection: Push the plunger slowly and pause a few seconds before withdrawing the needle to prevent backflow

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Protein was eluted (50 mM Tris, pH 7.5, 1 M NaCl, 500 mM imidazole and 2 mM TCEP) and the His6-Protein A tag was cleaved overnight at 4 °C using 2 (w/w) % of 1 mg/ml recombinant His-tagged TEV protease