Figure 1. Western blot analysis of SLC6A4 using anti-SLC6A4 antibody (PB9438). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human HepG whole cell lysates, Lane 2: human Caco-2 whole cell lysates, Lane 3: human SW620 whole cell lysates, Lane 4: human COLO-320 whole cell lysates. Lane 5: human K562 whole cell lysates, Lane 6: human U2OS whole cell lysates, Lane 7: human U-87MG whole cell lysates, Lane 8: rat heart tissue lysates, Lane 9: mouse heart tissue lysates, Lane 10: mouse SP20 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-SLC6A4 antigen affinity purified polyclonal antibody (Catalog # PB9438) at 0.5 microg/mL overnight at 4°C, 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SLC6A4 at approximately 90KD. The expected band size for SLC6A4 is at 70KD.