Journal: Protein Expression and Purification

Article Title: Expression, purification and characterization of recombinant severe acute respiratory syndrome coronavirus non-structural protein 1

doi: 10.1016/j.pep.2006.11.005

Figure Lengend Snippet: Coomassie stained SDS–PAGE analysis of SARS-CoV nsp1 at various stages of purification. (a) Gel displaying fractions from the initial IMAC purification (IMAC #1) and the second IMAC following cleavage of the dual affinity tag (IMAC #2). Lane 1, Mark12 molecular weight marker (Invitrogen); lane 2, total cell lysate; lane 3, lysate supernatant; lane 4, IMAC #1 flow-through; lane 5, IMAC #1 wash; lane 6, IMAC #1 HisGST-Snsp1 elution; lanes 7–9, IMAC #2 flow-through fractions containing SARS-CoV nsp1; lane 10, IMAC #2 elution of cleaved dual affinity tag. (b) Gel analysis of preparative SEC. Lane 1, Mark12 molecular weight marker (Invitrogen); lane 2, sample loaded onto SEC column; lanes 3–4, pooled SARS-CoV nsp1 eluted from SEC loaded onto the gel at 3 and 6 μg, respectively.

Article Snippet: The lysate supernatant was filtered through a 0.45 μM membrane, and then the clarified sample was loaded onto a 5 mL HisTrap immobilized metal affinity chromatography (IMAC) column (Amersham) equilibrated with binding buffer (50 mM Hepes, pH 7.5, 250 mM NaCl, 1 mM β-mercaptoethanol and 15 mM imidazole).

Techniques: Staining, SDS Page, Purification, Molecular Weight, Marker

Journal: Protein Expression and Purification

Article Title: Expression, purification and characterization of recombinant severe acute respiratory syndrome coronavirus non-structural protein 1

doi: 10.1016/j.pep.2006.11.005

Figure Lengend Snippet: Yield of recombinant SARS-CoV nsp1 purified from E. coli

Article Snippet: The lysate supernatant was filtered through a 0.45 μM membrane, and then the clarified sample was loaded onto a 5 mL HisTrap immobilized metal affinity chromatography (IMAC) column (Amersham) equilibrated with binding buffer (50 mM Hepes, pH 7.5, 250 mM NaCl, 1 mM β-mercaptoethanol and 15 mM imidazole).

Techniques: Recombinant, Purification