PMC:2724026 / 31866-32489 JSONTXT

{"project":"2_test","denotations":[{"id":"19362094-8757792-62520551","span":{"begin":70,"end":72},"obj":"8757792"}],"text":"Protein expression was carried out in Escherichia coli C41(DE3) cells:59 transformed cells were grown to an A600 of 0.4 – 0.6 at 37 °C before induction and growth overnight at 28 °C. The harvested cells were lysed by sonication and the protein purified from the soluble fraction, after centrifugation, by binding to Ni-NTA agarose (Qiagen). The bound protein was cleaved from the resin using thrombin (Sigma) and further purified using a Superdex G75 gel-filtration column (GE Healthcare). The purified protein was stored at 4 °C in 50 mM sodium phosphate, pH 7.0, 150 mM NaCl, 0.2 mM Tris(2-carboxyethyl)-phosphine (TCEP)."}