Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) is a rabbit monoclonal antibody that is used to detect HNF-4-alpha in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency
-Over 10 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
ChIC/CUT&RUN-seq | WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Expected | Tested | Expected | Expected | Tested |
Rat | Predicted | Expected | Expected | Expected | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 5 µg | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Mouse | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/2000 | Notes - |
Species Mouse | Dilution info 1/2000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species Rat | Dilution info 1/1000 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
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Transcriptional regulator which controls the expression of hepatic genes during the transition of endodermal cells to hepatic progenitor cells, facilitating the recruitment of RNA pol II to the promoters of target genes (PubMed:30597922). Activates the transcription of CYP2C38 (By similarity). Represses the CLOCK-BMAL1 transcriptional activity and is essential for circadian rhythm maintenance and period regulation in the liver and colon cells (PubMed:30530698).
HNF4, NR2A1, TCF14, HNF4A, Hepatocyte nuclear factor 4-alpha, HNF-4-alpha, Nuclear receptor subfamily 2 group A member 1, Transcription factor 14, Transcription factor HNF-4, TCF-14
Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) is a rabbit monoclonal antibody that is used to detect HNF-4-alpha in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency
-Over 10 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Mouse species are recommended based on IHC result. This antibody is unsuitable to be used in WB on mouse.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
HNF-4-alpha also known as hepatocyte nuclear factor 4-alpha (HNF4A) is a nuclear protein that functions as a transcription factor. It plays an important role in regulating gene expression in the liver kidney and intestine. HNF-4-alpha has a molecular mass of about 54 kDa and is expressed in tissues where it maintains cellular differentiation and metabolic homeostasis. In cellular assays such as ELISA and CHIP detection of HNF-4-alpha provides insights into its expression levels and activity in various biological contexts.
HNF-4-alpha influences the expression of genes involved in liver and pancreatic function. It acts as a transcriptional regulator that controls fatty acid oxidation gluconeogenesis and insulin secretion. HNF-4-alpha interacts with other proteins to form transcriptional complexes that fine-tune the expression of metabolic enzymes and transport proteins. This protein is key to ensuring that metabolic genes are expressed in a coordinated manner which is essential for maintaining normal metabolic processes.
HNF-4-alpha participates in the insulin signaling and lipid metabolism pathways. Through these pathways it interacts with other transcription factors such as HNF-1-alpha and PPAR-alpha which coordinate the regulation of genes involved in glucose metabolism and fatty acid oxidation. HNF-4-alpha plays a critical role by modulating the expression of enzymes like gluconeogenic enzymes in response to metabolic needs which connects it to key metabolic pathways influencing overall energy homeostasis.
HNF-4-alpha is linked to maturity-onset diabetes of the young (MODY1) and certain types of liver diseases. Mutations or dysregulation in HNF-4-alpha can affect the expression of genes essential for insulin production and glucose metabolism leading to disorders like diabetes. Additionally HNF-4-alpha interacts with other proteins and transcription factors implicated in these conditions such as HNF-1-alpha. Understanding the role of HNF-4-alpha in these diseases can provide insights into potential therapeutic targets and intervention strategies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Blocking/Dilution buffer: 5% NFDM/TBST.
Based on sequence analysis of the immunogen, ab201460 shares 72% homology with HNF4 gamma. The levels of XR were tested in the accompanying recombinant protein, WB experiment.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution
All lanes: Full length mouse HNF-4-gamma recombinant protein at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Exposure time: 3min
Intracellular Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) labelling CDKN2A/p16INK4a with purified ab201460 at 1/1000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton-X100 permeabilized HT-29 (Human colorectal adenocarcinoma cells) cells labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/500 dilution (green).
Confocal image showing nuclear staining on HT-29 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution (red).
-ve control 1: ab201460 at 1/2000 dilution followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/500 dilution.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton-X100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/500 dilution (green).
Confocal image showing nuclear staining on HT-29 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution (red).
-ve control 1: ab201460 at 1/2000 dilution followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/500 dilution.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/10000 dilution
All lanes: HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution
All lanes: Caco-2 (Human colorectal adenocarcinoma cells) whole cell lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 15s
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution
Lane 1: Human colon lysate at 10 µg
Lane 2: Human fetal kidney lysate at 10 µg
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 1min
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on Human colon tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on Human liver tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on mouse liver tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution.
Nuclear staining on rat colon tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (Human liver hepatocellular carcinoma cell line) cells and 5 µg of ab201460 [EPR16885-99)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.
Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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