NSDHL Antibody / NADP-dependent steroid dehydrogenase-like protein

Western blot analysis of NSDHL using anti-NSDHL antibody. Lane 1: human whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: human RT4 whole cell lysates, Lane 4: human CACO-2 whole cell 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-NSDHL antibody at 1:500 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. NSDHL antibody detects a single band migrating at ~38 kDa across the indicated cell and tissue lysates. Although the theoretical molecular weight is ~42 kDa, NSDHL is an ER-associated membrane protein that typically migrates faster on SDS-PAGE due to signal peptide cleavage and altered SDS binding. The observed size is consistent with published reports of the mature NSDHL form.