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Knockout Tested Rabbit Recombinant Monoclonal HNF-4-alpha antibody. Suitable for ChIC/CUT&RUN-seq, IP, ChIP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 2 publications.


Images

ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (AB200142), expandable thumbnail
  • Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (AB200142), expandable thumbnail
  • Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (AB200142), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (AB200142), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (AB200142), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Abcam Recommends

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ChIC/CUT&RUN-seqIPChIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Tested

Tested
Tested

Species
Human
Dilution info
5 µg
Notes

-

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Tested
Tested

Species
Human
Dilution info
5 µg chromatin for 25.00000 µg chromatin
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100
Notes

-

Tested
Tested

Species
Human
Dilution info
1/600
Notes

-

Associated Products

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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. Suitable for ChIC/CUT&RUN-seq, IP, ChIP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 2 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR19265-130
Purification technique
Affinity purification Protein A
Concentration
Loading...

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:
Terms & Conditions.

8 product images

  • ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142), expandable thumbnail

    ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

    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 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).

  • Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142), expandable thumbnail

    Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

    Exposure times: Lanes 1 and 2: 3 minutes; Lanes 3 and 4: 15 seconds; Lane 5: 3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142) at 1/2000 dilution

    Lane 1: Human colon lysate at 20 µg

    Lane 2: Human fetal liver lysate at 20 µg

    Lane 3: HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

    Lane 4: Caco-2 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

    Lane 5: SW480 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/2000 dilution

    Developed using the ECL technique.

    Predicted band size: 53 kDa

    Observed band size: 53 kDa

  • Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142), expandable thumbnail

    Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

    HNF-4-alpha was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab200142 at 1/30 dilution. Western blot was performed from the immunoprecipitate using 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: ab200142 IP in HepG2 whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab200142 in HepG2 whole cell lysate (-).
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

    All lanes: Immunoprecipitation - 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 (ab200142), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

    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 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SW480 (human colorectal adenocarcinoma cell line) cells labeling HNF-4-alpha with 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.

  • Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142), expandable thumbnail

    Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

    Western blot: Anti-HNF4A antibody [EPR19265-130] (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, 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 (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

    Secondary

    All 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 (ab200142), expandable thumbnail

    ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

    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 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.

  • Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

    Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cell line labeling HNF-4-alpha with 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.

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Product protocols

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