ZYX Antibody / Zyxin

Western Blot: 0.25-0.5 µg/mL, Immunohistochemistry: 2-5 µg/mL, Immunocytochemistry: 5 µg/mL, Immunofluorescence: 5 µg/mL, Immunoprecipitation: 2-4ug/500ug of lysate, Flow cytometry: 1-3ug/million cells

Western blot analysis of Zyxin using anti-Zyxin antibody. Lane 1: human MCF-7 whole cell lysates, Lane 2: human PC-3 whole cell lysates, Lane 3: human SIHA whole cell lysates, Lane 4: rat lung tissue lysates, Lane 5: mouse lung tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Zyxin antibody at 1:1000 overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:500 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. Zyxin (ZYX, ~61 kDa predicted) was detected at ~75-80 kDa, consistent with its phosphorylated and isoform-dependent mobility reported in the literature. Lower bands (~35-45 kDa) likely represent proteolytic fragments generated during turnover or stress responses.