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. 2016 Aug 20;8:127–138. doi: 10.1016/j.bbrep.2016.08.016

Fig. 2.

Fig. 2.

Purification of LdGcs. (A) 10% SDS PAGE of GcsF purification samples Lane 1–5 correspond to unstained protein marker; cell lysate loaded on NiNTA column; flow through of NiNTA column; wash sample conataining 10 mM imidazole; wash sample containing 50 mM imidazole; lane 6–7 elution sample containing NiNTA purified Gcs; right panel anti-His western blot of elution sample showing bands are degradation products of Gcs. (B) Size exclusion chromatography profile of Gcs with elution volume 15.8 ml corresponding to reference protein conalbumin (75 kDa) showing molecular weight corresponding to monomer. Peak at 8.8 corresponds to molecular weight above 440 corresponding to higher order oligomer. (C) 10% SDS PAGE showing M, unstained standards; lanes 1 and 2, over-expression of GcsT (DE3) pre-induction (lane 1) and post-induction (lane 2), with 1 mM IPTG. Lane 3, Western immunoblotting of over-expressed sample with anti-His antibody. Lane 4–5 purified GCST on 10% SDS PAGE (D) 10% SDS PAGE showing M unstained standards; lanes 1 and 2, over-expression of pET28a-GcsC (DE3) pre-induction and post-induction with 1 mM IPTG, M, prestained protein molecular weight marker; lane 3, Purified GcsC at expected size 45 kDa.