Journal:

Article Title: Species-Specific Receptor Recognition by a Minor-Group Human Rhinovirus (HRV): HRV Serotype 1A Distinguishes between the Murine and the Human Low-Density Lipoprotein Receptor

doi: 10.1128/JVI.76.14.6957-6965.2002

Figure Lengend Snippet: Western blot analysis of membrane proteins from HeLa and M cell lines grown under normal conditions or under conditions of up-regulation of LDLR expression (↑). Membrane proteins extracted from approximately 106 cells were separated on SDS-polyacrylamide gels with 8% (A; C, top), 4.5 to 18% (B), and 4.5 to 10% (C, bottom) acrylamide under nonreducing conditions and electrophoretically transferred to PVDF membranes. After blocking with 2% Tween 20 in PBS, LDLR was revealed with antibodies against human LDLR followed by enzyme-conjugated secondary antibodies. LRP was detected with Myc-tagged scFv7 followed by mouse anti-Myc antibody 9E10 and HRP-conjugated goat anti-mouse antibody. Note that in the bottom gel in panel C, twice the amount of membrane proteins was used for HeLa cells compared to M1 cells, and the exposure times for detection using chemiluminescence were 1.5 min for HeLa cells and 10 s for M1 cells.

Article Snippet: The Myc-tagged human single-chain antibody scFv7 (0.51 mg/ml), which is directed against the chicken homologue of mammalian VLDLR and cross-reacts with mouse and human LRP ( 16 ), was used at a 1:2,000 dilution; as a secondary antibody, the mouse anti-Myc antibody 9E10 was employed at 1 μg/ml followed by HRP-conjugated goat anti-mouse antibody (Bio-Rad Laboratories, Hercules, Calif.).

Techniques: Western Blot, Expressing, Blocking Assay