Name: Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade
SKU: ab181604
Rating: 5 (4 reviews)

Anti-HNF-4-alpha antibody [EPR16885] ab181604 is a rabbit monoclonal antibody that is used in HNF-4-alpha western blotting and IHC. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR16885 is cited in over 50 publications

Images

ChIP - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (AB181604), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ChIC/CUT&RUN-seq	IP	ChIP	WB	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Expected	Expected	Tested	Tested
Rat	Expected	Expected	Expected	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 5 µg	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/70	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 2 µg chromatin for 25.00000 µg chromatin	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

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 Rat	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.

Associated Products

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Target data

See full target information HNF4A

Function

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).

Alternative names

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR16885
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

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:
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13 product images

ChIP - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Chromatin was prepared from HepG2 (Human liver hepatocellular carcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab181604 (blue), and 20μl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
ABCC6_NC1 is negative control
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour before being incubated with ab181604 (rabbit monoclonal [EPR16885] to HNF-4-alpha; dilution 1:10000) and loading control ab18058 (mouse monoclonal [SPM227] to Vinculin; dilution 1:10000) at 4°C overnight. Antibody binding was detected with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 (Goat anti-Rabbit IgG H&L (IRDye® 800CW); green; dilution 1:20000) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 (Goat anti-Mouse IgG H&L (IRDye® 680RD), red; dilution 1:20000) for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/10000 dilution
Lane 1: HepG2 whole cell lysate at 20 µg
Lane 2: HEK293 whole cell lysate at 20 µg
Lane 3: Mouse liver lysate at 20 µg
Secondary
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Predicted band size: 53 kDa
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution
All lanes: Full length mouse HNF-4-gamma recombinant protein at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 45 kDa, 53 kDa
Observed band size: 38 kDa, 39 kDa
Exposure time: 3min
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/10000 dilution
All lanes: HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution
All lanes: SW480 (Human colon adenocarcinoma cell line) whole cell lysate at 10 µg
Secondary
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: 5s
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution
Lane 1: Human fetal liver lysate at 10 µg
Lane 2: Human colon lysate at 10 µg
Lane 3: Human fetal kidney lysate at 10 µg
Secondary
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
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
This antibody can recognize 2 isoforms in mouse and rat. The predicted MW are 53KDa and 52 KDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution
Lane 1: Mouse liver lysate at 10 µg
Lane 2: Rat liver lysate at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Exposure time: 15s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HNF-4-alpha with ab181604 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. Nucleus staining on Human liver 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.
Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
HNF-4 alpha was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell extract with ab181604 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab181604 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: HepG2 whole cell extract 10 μg (Input).
Lane 2: ab181604 IP in HepG2 whole cell extract.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab181604 in HepG2 whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 5s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HNF-4-alpha with ab181604 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 epithelial cells of Human colon 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HNF-4-alpha with ab181604 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 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling HNF-4-alpha with ab181604 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 epithelial cells of rat colon 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 sequencing - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
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 ab181604 [EPR16885]. 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 downloaded here.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.