D-dopachrome decarboxylase DDT Rabbit Polyclonal Antibody

Western blot analysis of DDT using anti-DDT antibody (orb76359, Left) and anti-DDT antibody (orb27669, Right). 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 30 ug of sample under reducing conditions. Lane 1: human hepatocellular carcinoma tumor tissue (HCCT) lysates, Lane 2: rat liver tissue lysates, Lane 3: mouse liver tissue 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-DDT antigen affinity purified polyclonal antibody (Catalog # orb76359) and rabbit anti-DDT antigen affinity purified polyclonal antibody (Catalog # orb27669) at 0.5 μg/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 # orb90503) with Tanon 5200 system. A specific band was detected for DDT at approximately 13-15 kDa. The expected band size for DDT is at 13 kDa.

IHC analysis of DDT using anti-DDT antibody (orb76359). DDT was detected in paraffin-embedded section of human placenta tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-DDT Antibody (orb76359) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.