Anti EGFR  

Western blot using Rockland's Affinity Purified anti-EGFR antibody shows detection of a band at ~170 kDa corresponding to human EGFR (arrowhead). Lane 1: unstimulated A431 whole cell lysate (p/n W09-000-361). Lane 2: EGF (50 ng/ml for 15 min) stimulated A431 whole cell lysates (p/n W09-000-362). Approximately 30μg of lysate was separated on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose. After blocking the membrane was probed with the primary antibody diluted to 1:1,000. Reaction occurred overnight at 4° C followed by washes and reaction with a 1:10,000 dilution of IRDye800 conjugated Gt-a-Rabbit IgG [H&L] MX (p/n 611-132-122) for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red). IRDye800 fluorescence image was captured using the OdysseyR Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.