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. 2022 Oct 10;79(1):284–294. doi: 10.1002/ps.7198

Figure 1.

Figure 1

(a) Schematic of constructs encoding recombinant HxTx‐Hv1h and HxTx‐Hv1h/GNA produced in the yeast P. pastoris showing predicted molecular masses; tag denotes the presence of a six‐residue histidine sequence that allows protein purification by nickel affinity chromatography and detection by Western blotting. (b) Separation of purified proteins by SDS‐PAGE gel stained for total protein: lanes 1 and 2 are HxTx‐Hv1h (c. 1 and 3 μg, respectively), lanes 3 and 4 are HxTx‐Hv1h/GNA (c. 2 and 3 μg, respectively), lanes 5 and 6 are recombinant GNA (c. 3 and 5 μg, respectively) and lane 7 is Sigma GNA standard (2 μg). (c) Western analysis of recombinant proteins using anti‐His [lanes 1–5 are as for (b); loading c. 50 and 100 ng for HxTx‐Hv1h and HxTx‐Hv1h/GNA and 50 ng for GNA] and anti‐GNA (lanes 1 and 2 are, respectively, 50 and 100 ng HxTx‐Hv1h/GNA, lane 3 is Sigma GNA standard 50 ng) antibodies. Location of mass markers run on the same gel are depicted. (d) Separation of nondenatured and denatured proteins by native PAGE, gel‐stained for total protein (15 μg loaded in each lane). Lanes 1 and 4 are HxTx‐Hv1h/GNA, lanes 2 and 5 are HxTx‐Hv1h, and lanes 3 and 6 are recombinant GNA.