WB
Western blot analysis of Alpha-Actinin using anti- Alpha-Actinin antibody. 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: mouse heart tissue lysates,
Lane 2: mouse skeletal muscle tissue lysates,
Lane 3: rat heart tissue lysates,
Lane 4: rat skeletal muscle 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 mouse anti- Alpha-Actinin antigen affinity purified monoclonal antibody at 4 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a Biotin Conjugated goat anti-mouse IgG 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 with Tanon 5200 system.
IHC
IHC analysis of Alpha-Actinin using anti- Alpha-Actinin antibody.Alpha-Actinin was detected in paraffin-embedded section of mouse lung 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 1.5μg/ml mouse anti- Alpha-Actinin Antibody 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) with DAB as the chromogen.
IHC
IHC analysis of Alpha-Actinin using anti- Alpha-Actinin antibody.Alpha-Actinin was detected in paraffin-embedded section of human mammary cancer 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 1.5μg/ml mouse anti- Alpha-Actinin Antibody 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) with DAB as the chromogen.