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Rabbit Recombinant Monoclonal HNF-4-alpha antibody. Suitable for ChIC/CUT&RUN-seq, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 18 publications.


Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
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.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ChIC/CUT&RUN-seqWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Expected
Tested
Rat
Predicted
Expected
Expected
Expected
Predicted

Tested
Tested

Species

Human

Dilution info

5 µg

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Rat

Dilution info

-

Notes

-

Tested
Tested

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

-

Expected
Expected

Species

Rat

Dilution info

1/1000

Notes

-

Tested
Tested

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.

Expected
Expected

Species

Rat

Dilution info

1/2000

Notes

-

Species

Mouse

Dilution info

1/2000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

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.

Expected
Expected

Species

Mouse

Dilution info

1/1000

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Species

Rat

Dilution info

1/1000

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

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.

Predicted
Predicted

Species

Rat

Dilution info

-

Notes

-

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-ARNTL/BMAL1 transcriptional activity and is essential for circadian rhythm maintenance and period regulation in the liver and colon cells (PubMed:30530698).

Alternative names

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Rabbit Recombinant Monoclonal HNF-4-alpha antibody. Suitable for ChIC/CUT&RUN-seq, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 18 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Clone number

EPR16885-99

Purification technique

Affinity purification Protein A

Specificity

Mouse species are recommended based on IHC result. This antibody is unsuitable to be used in WB on mouse.

Concentration
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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.

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.

12 product images

  • Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460), expandable thumbnail

    Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 53 kDa

    Exposure time: 3min

  • Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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 (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 ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution (red).
    -ve control 1: ab201460 at 1/2000 dilution followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution.
    -ve control 2: ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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 (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 ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution (red).
    -ve control 1: ab201460 at 1/2000 dilution followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution.
    -ve control 2: ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution.

  • Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460), expandable thumbnail

    Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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

    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 [EPR16885-99] (ab201460), expandable thumbnail

    Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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

    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: 15s

  • Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460), expandable thumbnail

    Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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) (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) (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 [EPR16885-99] (ab201460), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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) (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) (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 [EPR16885-99] (ab201460), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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) (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) (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 [EPR16885-99] (ab201460), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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) (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) (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 [EPR16885-99] (ab201460), expandable thumbnail

    ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

    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 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.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

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