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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 7. PDZ Domain and PDZ Domain-binding Beta Hairpin (PDZbh) Affinity Tags.

Figure 7

All samples and Precision Plus Protein All Blue Standards were run on a 4-12% SDS-PAGE gradient gel and stained with Gel Code Blue.

(A) Soluble lysates after heterologous expression of MBP, GST, a PDZ2 domain, MBP fusion proteins containing an N-terminal or C-terminal PDZ2 Tag, and an MBP-GST fusion protein containing an internal PDZ2 Domain Tag.

(B) Peptide eluates from purifications of the proteins shown in (A) on HaloTag-HaloLink Affinity Resins containing one (1x PDZbh) or two (2x PDZbh) PDZbh Domains, and on PDZ2 Domain Peptide Ligand-NHS-Agarose resin, prepared as described in Methods. Proteins were eluted with 400 μg/ml SIESDV peptide as described in Methods.

(C) Soluble lysates after heterologous expression of MBP; GST; single and tandem PDZbh domains; and MBP, GST, and MBP-GST fusion proteins containing zero, one or two N-terminal, C-terminal or internal PDZbh domains.

(D) Peptide eluates from purification of the proteins shown in (C) on PDZ2-HaloTag-HaloLink domain resin eluted with 400 μg/ml SIESDV peptide, as described in Methods.