Figure 2. IHC analysis of Vinculin using anti-Vinculin antibody (MA1103). Vinculin was detected in frozen 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 1ug/ml rabbit anti-Vinculin Antibody (MA1103) overnight at 4°C. Biotinylated goat anti- mouse 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 # SA1021) with DAB as the chromogen.

Figure 3. Western blot analysis of Vinculin using anti-Vinculin antibody (MA1103). 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 Hela, whole cell lysates, . Lane 2: human HepG2, whole cell lysates, . Lane 3: monkey COS-7 whole cell lysates, . Lane 4: human U-87MG whole cell lysates,. Lane 5: rat PC-12 whole cell lysates, . Lane 6: rat RH35 whole cell lysates, . Lane 7: mouse HEPA1-6 whole cell lysates, . Lane 8: mouse NIH3T3 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 mouse anti-Vinculin antigen affinity purified monoclonal antibody (Catalog # MA1103) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti- mouse 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 # EK1001) with Tanon 5200 system. A specific band was detected for Vinculin at approximately 124KD. The expected band size for Vinculin is at 124KD.