antibody

Anti-HNF-4-alpha/HNF4A Picoband Antibody

PB9215

100 ug/vial

polyclonal

domestic rabbit

nonconjugated

human, mouse, rat

western blot, immunocytochemistry, flow cytometry, immunohistochemistry - paraffin section, immunohistochemistry - frozen section

Enabled

Anti-HNF-4-alpha/HNF4A Picoband Antibody

Primary Antibodies, Polyclonal Antibodies, IHC ICC IF Antibodies

HNF4A

Unconjugated / $280 APC / $330 APC-Cy7 / $330 FITC / $330 PE / $330 PE-Cy5 / $330 PE-Cy7 / $330 30ug sample size / $99 100ug / $280 100ug+Free HRP Secondary BA1054 / $280 100ug+Free Biotin Secondary BA1003 / $280

Polyclonal

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

No

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Polyclonal antibody for NR2A1/HNF4A detection. Host: Rabbit.Size: 100ug/vial. Tested applications: IHC-P. Reactive species: Human. NR2A1/HNF4A information: Molecular Weight: 52785 MW; Subcellular Localization: Nucleus.

Rabbit IgG polyclonal antibody for Hepatocyte nuclear factor 4-alpha(HNF4A) detection. Tested with WB, IHC-P, IHC-F, ICC/IF, FCM in Human;Mouse;Rat.

100 ug/vial

30ug for $99, contact us for details

P41235

Rabbit

E.coli-derived human HNF-4-alpha recombinant protein (Position: Q164-I474). Human HNF-4-alpha shares 95% and 96% amino acid (aa) sequences identity with mouse and rat HNF-4-alpha, respectively.

Lyophilized

Immunogen affinity purified.

At -20 C for one year. After reconstitution, at 4 C for one month. It can also be aliquotted and stored frozen at -20 C for a longer time.Avoid repeated freezing and thawing.

No cross reactivity with other proteins

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Western blot, 0.1-0.5ug/ml, Human, Rat. Immunohistochemistry(Paraffin-embedded Section), 0.5-1ug/ml, Human, Mouse, Rat, By Heat. Immunohistochemistry(Frozen Section), 0.5-1ug/ml, Mouse, Rat, -. Immunocytochemistry/Immunofluorescence, 2ug/ml, Human. Flow Cytometry, 1-3ug/1x10 6 cells, Human

Flow Cytometry, IF, IHC-P, IHC-F, ICC, WB

Human, Mouse, Rat

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. lane 1: Recombinant Human HNF-4-alpha Protein 0.5ng. 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-HNF-4-alpha antigen affinity purified polyclonal antibody (Catalog # PB9215) 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-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 38KD. The expected band size for HNF-4-alpha is at 38KD. Figure 2. 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: Rat Liver Tissue Lysate, . Lane 2: Rat Kidney Tissue Lysate, . Lane 3: RH35 Whole Cell Lysate, . Lane 4: HELA Whole Cell Lysate, . Lane 5: HEPG2 Whole Cell Lysate. 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-HNF-4-alpha antigen affinity purified polyclonal antibody (Catalog # PB9215) 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-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 3. IHC analysis of HNF-4-alpha using anti-HNF-4-alpha antibody (PB9215). HNF-4-alpha was detected in paraffin-embedded section of Mouse Kidney Tissue. 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-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 4. IHC analysis of HNF-4-alpha using anti-HNF-4-alpha antibody (PB9215). HNF-4-alpha was detected in paraffin-embedded section of Rat Kidney Tissue. 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-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 5. 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 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-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 6. IHC analysis of HNF-4-alpha using anti-HNF-4-alpha antibody (PB9215). HNF-4-alpha was detected in frozen section of Rat Kindey Tissue. 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-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 7. IHC analysis of HNF-4-alpha using anti-HNF-4-alpha antibody (PB9215). HNF-4-alpha was detected in frozen section of Mouse Liver Tissue. 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-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 tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/mL rabbit anti- HNF-4-alpha Antibody (PB9215) overnight at 4°C. DyLight®488 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 (PB9215,1ug/1x10 6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10ug/1x10 6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x10 6 ) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Hepatocyte nuclear factor 4 alpha (HNF4A), also known as NR2A1, is a nuclear receptor that in humans is encoded by the HNF4A gene. It is mapped to 20q13.12. HNF4A is a nuclear transcription factor that binds DNA as a homodimer. The encoded protein controls the expression of several genes, including hepatocyte nuclear factor 1 alpha, a transcription factor which regulates the expression of several hepatic genes. This gene plays a role in development of the liver, kidney, and intestines. HNF4A is required for the PXR and CAR-mediated transcriptional activation of CYP3A4. This gene also plays a pivotal role in the expression and synthesis of SHBG, an important glycoprotein made primarily in the liver, which in addition to lowering insulin-resistance also serves in reducing levels of free Oestrogen as-well as prolonging the half-life of Testosterone.

sc 249209 sc 249208

Hepatocyte nuclear factor 4-alpha;HNF-4-alpha;Nuclear receptor subfamily 2 group A member 1;Transcription factor 14;TCF-14;Transcription factor HNF-4;HNF4A;HNF4, NR2A1, TCF14;

Hepatocyte nuclear factor 4-alpha

52785 MW

Transcriptionally controlled transcription factor. Binds to DNA sites required for the transcription of alpha 1- antitrypsin, apolipoprotein CIII, transthyretin genes and HNF1- alpha. May be essential for development of the liver, kidney and intestine.

Nucleus.

Hepatocyte nuclear factor 4-alpha

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.

Belongs to the nuclear hormone receptor family. NR2 subfamily.

5/9/19 7:23