Knockout Tested Rabbit Recombinant Monoclonal HNF-4-alpha antibody. Carrier free. Suitable for ChIC/CUT&RUN-seq, IP, ChIP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.
Images
![ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--chip-img8122.jpg/_jcr_content/renditions/webp-475.webp "ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487)")
![Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--immunoprecipitation-img5142.jpg/_jcr_content/renditions/webp-475.webp "Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487)")
![Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--immunocytochemistry-immunofluorescence-img24151.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487)")
![Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--immunocytochemistry-immunofluorescence-img71463.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487)")
![Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--western-blot-img435359.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (AB226487)")
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| ChIC/CUT&RUN-seq | IP | ChIP | WB | ICC/IF | Flow Cyt (Intra) | |
|---|---|---|---|---|---|---|
| Human | Tested | Tested | Tested | Tested | Tested | Tested |
ChIC/CUT&RUN-seq
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
ChIP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Associated Products
Select an associated product type
6 products for Alternative Product
2 products for Alternative Version
Target data
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
Recommended products
Knockout Tested Rabbit Recombinant Monoclonal HNF-4-alpha antibody. Carrier free. Suitable for ChIC/CUT&RUN-seq, IP, ChIP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- EPR19265-130
- Purification technique
- Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- Do Not Freeze
Notes
ab226487 is the carrier-free version of Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142.
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:
- - 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.
Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
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.
Not recommended
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.
7 product images
![ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--chip-img8122.jpg "ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
Chromatin was prepared from HepG2 (human hepatocellular carcinoma epithelial cell) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 5 µg of Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 (blue), and 20 µl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach).
ChIP was performed according to the literature (PMID: 18850323).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142).
![Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--immunoprecipitation-img5142.jpg "Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
HNF-4-alpha was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) 10 μg (Input).
Lane 2: Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 IP in HepG2 whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 in HepG2 whole cell lysate (-).
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142).
All lanes: Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142)
Developed using the ECL technique.
Predicted band size: 53 kDa
Observed band size: 53 kDa
![Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--immunocytochemistry-immunofluorescence-img24151.jpg "Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells labeling HNF-4-alpha with Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing nuclear staining on HepG2 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142).
![Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--immunocytochemistry-immunofluorescence-img71463.jpg "Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SW480 (human colorectal adenocarcinoma cell line) cells labeling HNF-4-alpha with Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing nuclear staining on SW480 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142).
![Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--western-blot-img435359.jpg "Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
This data was developed using the same antibody clone in a different buffer formulation (abAB200142).
Western blot: Anti-HNF4A antibody [EPR19265-130] (Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 was shown to bind specifically to HNF4A. A band was observed at 50-55 kDa in wild-type A549 cell lysates with no signal observed at this size in HNF4A knockout cell line. To generate this image, wild-type and HNF4A knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: HNF4A knockout A549 cell lysate at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: HEK-293 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 50 kDa, 55 kDa
![ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--chic-cut-run-sequencing-img420837.jpg "ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 HepG2 (Human liver hepatocellular carcinoma cell line) cells and 5 µg of Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 [EPR19265-130]. 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142).![Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--flow-cytometry-intracellular-img414065.jpg "Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cell line labeling HNF-4-alpha with Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142 at 1/600 (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade ab200142).
![ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--chic-cut-run-sequencing-img420837.jpg/_jcr_content/renditions/webp-475.webp "ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
![Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487), expandable thumbnail](https://content.abcam.com/products/images/hnf-4-alpha-antibody-epr19265-130-chip-grade-bsa-and-azide-free-ab226487--flow-cytometry-intracellular-img414065.jpg/_jcr_content/renditions/webp-475.webp "Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
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