After separation by SDS-PAGE, the proteins were transferred onto PVDF membranes. Membranes were blocked with 5% (w/v) dried skimmed milk in TBS (25 mM Tris-HCl pH 7.5, 150 mM NaCl) and incubated with primary antibodies followed by IRDye fluorescently labelled secondary antibodies (LI-COR). Membranes were scanned with an Odyssey near-infrared imager (LI-COR). Primary antibodies against hamster BiP (chicken anti-BiP, 1:2000;48 (link)), actin (mouse anti-actin, 1:2000; Abcam, cat. # AB3280), MANF (chicken anti-MANF, 1:1000; see below), FLAG-M1 (mouse anti-FLAG-M1, 1:1000; Sigma, cat. # F3040), and A1AT (mouse anti-A1AT monoclonal, 1:5000; Abcam, cat. # AB9399) were used.
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