Anti-Alpha Tubulin - Western Blot. Western blot of Affinity Purified anti-alpha Tubulin antibody shows detection of a predominant band at ~50 kD corresponding to alpha tubulin (arrowhead). Lanes correspond to whole cell lysates from mouse brain (lane 1), rat brain (lane 2), A431 cells (lane 3), Jurkat cells (lane 4) and HeLa cells (lane 5). Approximately 35 ug of each lysate was separated on a 4-20% Tris-HEPES gel by SDS-PAGE and transferred onto nitrocellulose. After blocking the membrane was probed with the primary antibody diluted to 1:1200. Reaction occurred 2h at room temperature followed by washes and reaction with a 1:10000 dilution of IRDye 800 conjugated Gt-a-Rb IgG [H&L] MX10 (for 45 min at room temperature. IRDye 800 fluorescence image was captured using the Odyssey Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.