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. Author manuscript; available in PMC: 2015 Jun 1.
Published in final edited form as: Protein Expr Purif. 2014 Mar 6;98:46–62. doi: 10.1016/j.pep.2014.02.015

Figure 8. Purification of Proteins Using PDZ Domain and PDZ Peptide Ligand Affinity Tags.

Figure 8

DasherGFP, LacZ and CAT were heterologously expressed with PDZ2 Domain or PDZ2 Peptide Ligand Tags and purified with the cognate Affinity Resin. For each protein, the soluble fraction (S) from the heterologous expression and the peptide eluate (E) after the purification were run on a 4-12% SDS-PAGE gradient gel with Precision Plus Protein All Blue Standards and stained with Gel Code Blue. (A) DasherGFP, LacZ and CAT were tagged at the C-terminus with the PDZ2 Domain Peptide Ligand, purified on PDZ2 Domain-NHS-Agarose Affinity Resin, and eluted with 400 μg/ml SIESDV peptide. (B) The three proteins were tagged at the N-terminus with a PDZ2 Domain, purified on PDZ2 Domain Peptide Ligand-NHS-Agarose resin, and eluted with 400 μg/ml SIESDV peptide. (C) The three proteins were tagged at the N-terminus with a PDZbh Domain or (D) two tandem PDZbh Domains, purified on PDZ2 Domain-NHS-Agarose Affinity Resin, and eluted with 400 μg/ml SIESDV peptide.