Anti-HNF-4-alpha/HNF4A Antibod

[list_product_images]Figure 1. Western blot analysis of HNF-4-alpha using anti-HNF-4-alpha antibody (PB9215).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: HEPG2 Whole Cell LysateAfter 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-HNF-4-alpha antigen affinity purified polyclonal antibody (Catalog # PB9215) 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 # EK1002) with Tanon 5200 system. A specific band was detected for HNF-4-alpha at approximately 53KD. The expected band size for HNF-4-alpha is at 53KD.|Figure 2. IHC analysis of HNF-4-alpha using anti- HNF-4-alpha antibody (PB9215).HNF-4-alpha was detected in paraffin-embedded section of human liver 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 1μg/ml rabbit anti- HNF-4-alpha Antibody (PB9215) overnight at 4°C. Peroxidase Conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.|Figure 3. IHC analysis of HNF-4-alpha using anti- HNF-4-alpha antibody (PB9215).HNF-4-alpha was detected in paraffin-embedded section of human intestinal 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 1μg/ml rabbit anti- HNF-4-alpha Antibody (PB9215) overnight at 4°C. Peroxidase Conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.|Figure 4. IHC analysis of HNF-4-alpha using anti- HNF-4-alpha antibody (PB9215).HNF-4-alpha was detected in paraffin-embedded section of mouse 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 1μg/ml rabbit anti- HNF-4-alpha Antibody (PB9215) overnight at 4°C. Peroxidase Conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.|Figure 5. IHC analysis of HNF-4-alpha using anti- HNF-4-alpha antibody (PB9215).HNF-4-alpha was detected in paraffin-embedded section of mouse liver 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 1μg/ml rabbit anti- HNF-4-alpha Antibody (PB9215) overnight at 4°C. Peroxidase Conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.|Figure 6. IHC analysis of HNF-4-alpha using anti- HNF-4-alpha antibody (PB9215).HNF-4-alpha was detected in paraffin-embedded section of rat liver 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 1μg/ml rabbit anti- HNF-4-alpha Antibody (PB9215) overnight at 4°C. Peroxidase Conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen. |Figure 7. IHC analysis of HNF-4-alpha using anti-HNF-4-alpha antibody (PB9215). HNF-4-alpha was detected in frozen section of mouse kindey tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HNF-4-alpha Antibody (PB9215) 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 # SA1022) with DAB as the chromogen.|Figure 8. IF analysis of HNF-4-alpha using anti- HNF-4-alpha antibody (PB9215).HNF-4-alpha was detected in immunocytochemical section of HEPG2 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti- HNF-4-alpha Antibody (PB9215) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.|Figure 9. Flow Cytometry analysis of HEPG2 cells using anti- HNF-4-alpha antibody (PB9215).Overlay histogram showing HEPG2 cells stained with PB9215 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HNF-4-alpha Antibody (PB9215,1μg/1x106 cells) for 30 min at 20°C. DyLight?488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.|Figure 10. IHC analysis of HNF-4-alpha using anti-HNF-4-alpha antibody (PB9215).HNF-4-alpha was detected in frozen section of rat liver tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-HNF-4-alpha Antibody (PB9215) 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 # SA1022) with DAB as the chromogen.[/list_product_images]