NME3 Antibody / Nucleoside diphosphate kinase 3 (FY13311)

Western blot analysis of NME3 using anti-NME3 antibody. Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human 293T whole cell lysates, Lane 2: human SIHA whole cell lysates, Lane 3: human MCF-7 whole cell lysates, Lane 4: human THP-1 whole cell lysates, Lane 5: rat kidney 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-NME3 antibody at 0.5 ug/ml 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 1:5000 for 1.5 hour at RT. The signal was developed using an ECL Plus Western Blotting Substrate. A predominant band is detected at an approximately 17 kDa in human cell lines, slightly below the predicted ~19 kDa size and consistent with the processed mature form of NME3. SiHa and THP-1 lysates also show a weaker band near 18 kDa, likely representing a partially processed or differently modified NME3 species. Rat kidney displays a band close to 19 kDa, consistent with species specific differences in processing or migration.