Western blot using Affinity Purified anti-beta Actin antibody shows detection of a predominant band at ~42 kDa corresponding to beta Actin (arrowhead) in various whole cell lysates. Lysates are as follows: human embryonic kidney 293 (lane 2), human lung carcinoma A549 (lane 3) and mouse brain (lane 4) all using the 800 nm channel (green). ~ 35 ug of each lysate was separated on a 4-20% Tris-glycine gel by SDSPAGE and transferred onto nitrocellulose. After blocking the membrane was probed with the primary antibody diluted to 1:1, 500. Incubation was overnight at 4°C followed by washes and reaction with a 1:10,000 dilution of IRDye (TM)800 conjugated Gt-a-Rabbit IgG [H&L] MX for 45 min at room temperature. Molecular weight markers are shown in lane 1 using the 700 nm channel (red). IRDye (TM)800 fluorescence image was captured using the Odyssey (R) Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.