BAK/BAK1 Antibody

Western blot analysis of BAK/BAK1 using anti-BAK/BAK1 antibody (orb738368). 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 PC-3 whole cell lysates, Lane 2: human HEK293 whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human CACO-2 whole cell lysates, Lane 5: human U20S whole cell lysates, Lane 6: human HEPG2 whole cell lysates, Lane 7: human HELA whole cell lysates, Lane 8: human A549 whole cell lysates, Lane 9: rat heart tissue lysates, Lane 10: rat lung tissue lysates, Lane 11: mouse heart tissue lysates, Lane 12: mouse lung 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-BAK/BAK1 antigen affinity purified polyclonal antibody (Catalog # orb738368) 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 1:5000 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 BAK/BAK1 at approximately 23KD. The expected band size for BAK/BAK1 is at 23KD.

IHC analysis of BAK/BAK1 using anti-BAK/BAK1 antibody (orb738368). BAK/BAK1 was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-BAK/BAK1 Antibody (orb738368) 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.

IHC analysis of BAK/BAK1 using anti-BAK/BAK1 antibody (orb738368). BAK/BAK1 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-BAK/BAK1 Antibody (orb738368) 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.

IF analysis of BAK/BAK1 using anti-BAK/BAK1 antibody (orb738368). BAK/BAK1 was detected in immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-BAK/BAK1 Antibody (orb738368) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.