WB analysis of transfected and non-transfected lysate using GTX48743 Ajuba antibody.
Lane 1 : Vector only
Lane 2 : Human Ajuba-RFP
Lane 3 : Mouse Ajuba-RFP
Lane 4 : Mock
Loading : 50 μg
Dilution : 1:700

Western blot using Affinity Purified anti-Ajuba antibody shows detection of a 57-kDa band consistent with the expected MW for Ajuba (arrowhead). Lanes correspond to 1) HeLa nuclear extract, and 2) HeLa, 3) A431, 4) Jurkat and 5) 293 whole cell lysates. Immunoprecipitation of Ajuba followed by western blotting may result in cleaner background staining. Approximately 5 μg of each preparation was run on a SDS-PAGE and transferred onto nitrocellulose followed by reaction with a 1:500 dilution of anti-Ajuba antibody. Detection occurred using a 1:5,000 dilution of HRP-labeled Donkey anti-Rabbit IgG for 1 hour at room temperature. A chemiluminescence system was used for signal detection (Roche) using a 60-sec exposure time.

Western blot using Affinity Purified anti-Ajuba antibody shows detection of Ajuba-RFP fusion protein in cell lysates (arrow-head). Lanes correspond to 1) vector only trans-fection, 2) human Ajuba-RFP, 3) mouse Ajuba-RFP, and 4) mock transfection. Approximately 50 μg of each lysate was loaded per lane for SDS-PAGE followed by transfer onto nitrocellulose and reaction with a 1:1,700 dilution of anti-Ajuba antibody.