Figure 1.

Purification of the human SPCA1a isoform by affinity chromatography. Gel stained for an SDS-PAGE (A), mass spectrometry analysis (B), and immunoblotting (C and D) of hSPCA1a fractions collected during affinity purification. The yeast membrane fractions (m), the flow-through after binding to Ni-NTA beads (F), the flow-through of each washing step (W1–3), each elution fraction (E1–5), and the beads (B) after elution were collected. Equal volumes of each fraction were separated via SDS-PAGE. Immunoblotting was performed with antibodies against SPCA1a (C) or the C-terminal His tag (D). Images are representative for a minimum of n = 3 experiments. L, ladder.