Journal: The Journal of Biological Chemistry
Article Title: Myeloid-derived growth factor is a resident endoplasmic reticulum protein
doi: 10.1074/jbc.AC118.002052
Figure Lengend Snippet: Eosinophil MYDGF. A, immunoblots of lysate from 2 × 105 unactivated (−) or IL5-activated (+) eosinophils per lane and 10 ng of recombinant, human MYDGF (rM) produced and purified from E. coli. Ponceau S stain is representative of total lysate protein; the most intense band centered at 13.4 kDa (*) corresponded to a band found in all immunoblot lanes regardless of whether goat anti-MYDGF, rabbit anti-MYDGF, or anti-P4HB primary antibodies were used. Mature MYDGF and P4HB were both detected in close proximity to their theoretical molecular masses of 15.8 and 55.3 kDa, respectively. All immunoblot images were taken with a 3-min exposure and the same image settings. B, human eosinophils, unactivated or activated with IL5, triple-stained with DAPI (blue), goat anti-human MYDGF or control IgGs (green), and rabbit anti-human MYDGF or control IgGs (pseudocolored red). Cells were imaged by confocal fluorescence and transmission microscopy. Images are of single 0.3-μm slices. Arrowheads, patch between nuclear lobes; arrows, periphery of nucleus; scale bar, 10 μm. Immunostaining and immunoblots are representative of experiments conducted on three or more separate occasions.
Article Snippet: Goat anti-human MYDGF, sold as anti-human SF20 (AF1147), and IL5 (205-IL) were from R&D Systems (Bio-Techne Corp., Minneapolis, MN).
Techniques: Western Blot, Recombinant, Produced, Purification, Staining, Control, Fluorescence, Transmission Assay, Microscopy, Immunostaining
