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Figure 2 | Proteome Science

Figure 2

From: Online monitoring of immunoaffinity-based depletion of high-abundance blood proteins by UV spectrophotometry using enhanced green fluorescence protein and FITC-labeled human serum albumin

Figure 2

Cloning, overexpression, and purification of EGFP. (A) Map of pET24a (+)-EGFP-His, drawn with vector NTI software (version 6.0, InforMax). The EGFP gene is designated by the gray arrow. (B) pET24a (+)-EGFP-His was transformed into BL21 RIL codon-plus (DE3) (Novagen) and overexpressed by IPTG induction. Molecular weight marker (MW), uninduced total cell lysate (UN), induced total cell lysate (IN), and induced supernatant (SPNT) were loaded on a 12% SDS-PAGE gel and visualized using Coomassie Brilliant Blue R250. (C) Purification was performed by Ni-NTA, and the flow-through fraction (FT), wash fraction (W), and eluted fraction (EL) were run on a 12% SDS-PAGE gel. The EGFP-His band is indicated with a black arrow.

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ISSN: 1477-5956