Ab62363

ConA agarose, HPLC grade acetonitrile (ACN) and formic acid, trifluoroacetic acid, ammonium bicarbonate, iodoacetamide (IAA), and dithiothreitol (DTT) were purchased from Sigma (St. Louis, MO, USA). Sequencing grade trypsin was purchased from Promega (Madison, WI, USA). The 4-plex iTRAQ regents were purchased from ABsciex (Framingham, MA, USA). A TripleTOF 5600 mass spectrometer from ABsciex and an HPLC system from Waters (Milford, MA, USA) were used.
For western blot, the primary antibodies for Alpha-1-antitrypsin (SERRINA1) (SERRINA1, ab9400), Ceruloplasmin (CP, ab51083), Transthyretin (TTR, ab9015), Apolipoprotein A-IV (APOA4, ab81616), Pro-epidermal growth factor (EGF, ab9695) and Prolactin-inducible protein (GCDFP15, ab62363) were purchased from Abcam (Cambridge, UK).

Whole-cell lysate and centrifuged conditioned media, consisting of 50 μg protein, were denatured, separated on SDS-PAGE gels and transferred to polyvinylidene fluoride membranes. After blocking in 5% milk in Tris-buffered saline–Tween, membranes were probed overnight at 4℃. Primary antibodies include AR (PG-21, 1:500 dilution; EMD Millipore), PSA (A0562, 1:1000 dilution; DAKO), FKBP5 (#8245, 1:1000 dilution; Cell Signaling), ZAG (sc-13585, 1:1000 dilution; Santa Cruz), GCDFP 15 corresponding to PIP (ab62363, 1:1000 dilution; abcam) and GAPDH (G8795, 1:10,000 dilution; Sigma-Aldrich). For secondary antibodies, IRDye 800 goat anti-rabbit and IRDye 680 goat anti-mouse (LI-COR, Lincoln, NE, USA) were used at 1:10,000. Bands were visualized and quantitated with the Odyssey® CLx Imaging System (LI-COR, Lincoln, NE, USA). Ponceau S staining of the membranes was used as loading control for western blots of conditioned media.