Name: Anti-HNF-4-alpha antibody [EPR16885-99]
Brand: RabMAb
SKU: ab201460
Rating: 4 (1 reviews)

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16885-99] to HNF-4-alpha
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIC/CUT&RUN-seq
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-HNF-4-alpha antibody [EPR16885-99]
See all HNF-4-alpha primary antibodies
Description

Rabbit monoclonal [EPR16885-99] to HNF-4-alpha
Host species

Rabbit
Specificity

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

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIC/CUT&RUN-seqmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HepG2 and Caco-2 whole cell lysates; Human colon and fetal kidney lysates. IHC-P: Human colon, Human liver, mouse liver and rat colon tissues. ICC/IF: HepG2 and HT-29 cells. ChIC/CUT&RUN-Seq: HepG2 cells.
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16885-99
Isotype

IgG
Research areas

Alternative Versions
- Anti-HNF-4-alpha antibody [EPR16885-99] - BSA and Azide free (ab231167)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human HNF-4-alpha protein (ab132090)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab201460 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB		1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/2000.
ChIC/CUT&RUN-seq		Use at an assay dependent concentration. 5 µg

Notes
Flow Cyt (Intra) 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/2000.
ChIC/CUT&RUN-seq Use at an assay dependent concentration. 5 µg

Function

Transcriptionally controlled transcription factor. Binds to DNA sites required for the transcription of alpha 1-antitrypsin, apolipoprotein CIII, transthyretin genes and HNF1-alpha. May be essential for development of the liver, kidney and intestine.
Involvement in disease

Defects in HNF4A are the cause of maturity-onset diabetes of the young type 1 (MODY1) [MIM:125850]; also symbolized MODY-1. MODY is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease.
Sequence similarities

Belongs to the nuclear hormone receptor family. NR2 subfamily.
Contains 1 nuclear receptor DNA-binding domain.
Post-translational
modifications

Phosphorylated on tyrosine residue(s); phosphorylation is important for its DNA-binding activity. Phosphorylation may directly or indirectly play a regulatory role in the subnuclear distribution.
Cellular localization

Nucleus.
Target information above from: UniProt accession P41235 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3172 Human
- Entrez Gene: 15378 Mouse
- Entrez Gene: 25735 Rat
- Omim: 600281 Human
- SwissProt: P41235 Human
- SwissProt: P49698 Mouse
- SwissProt: P22449 Rat
- Unigene: 116462 Human
- Unigene: 202383 Mouse
- Unigene: 12238 Rat
see all
Alternative names
- FLJ39654 antibody
- FRTS4 antibody
- Hepatic nuclear factor 4 alpha antibody
- Hepatocyte nuclear factor 4 alpha antibody
- Hepatocyte nuclear factor 4 antibody
- Hepatocyte nuclear factor 4-alpha antibody
- HNF 4 alpha antibody
- HNF 4 antibody
- HNF-4-alpha antibody
- HNF4 antibody
- HNF4A antibody
- HNF4A_HUMAN antibody
- HNF4a7 antibody
- HNF4a8 antibody
- HNF4a9 antibody
- Hnf4alpha antibody
- HNF4alpha10/11/12 antibody
- MODY 1 antibody
- MODY antibody
- MODY1 antibody
- NR2A1 antibody
- NR2A21 antibody
- Nuclear receptor subfamily 2 group A member 1 antibody
- OTTHUMP00000031060 antibody
- OTTHUMP00000031062 antibody
- TCF 14 antibody
- TCF antibody
- TCF-14 antibody
- TCF14 antibody
- Tcf4 antibody
- Transcription factor 14, hepatic nuclear factor antibody
- Transcription factor 14 antibody
- Transcription factor HNF 4 antibody
- Transcription factor HNF-4 antibody
- Transcription factor HNF4 antibody
see all

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/µL, 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 downloaded here.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution + Full length mouse HNF-4-gamma recombinant protein at 10 µg

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

Predicted band size: 53 kDa

Exposure time: 3 minutes

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.
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)

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)

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)

Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/10000 dilution + HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 10 µg

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

Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution + Caco-2 (Human colorectal adenocarcinoma cells) whole cell lysate at 10 µg

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

Predicted band size: 53 kDa
Observed band size: 53 kDa

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

All lanes : Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution

Lane 1 : Human colon lysate
Lane 2 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

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: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
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)

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)

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)

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

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab201460? Please let us know so that we can cite the reference in this datasheet.

ab201460 has been referenced in 14 publications.

Xu RC et al. A novel murine model of combined hepatocellular carcinoma and intrahepatic cholangiocarcinoma. J Transl Med 20:579 (2022). PubMed: 36494846
Lee Y et al. Hepatic stellate and endothelial cells maintain hematopoietic stem cells in the developing liver. J Exp Med 218:N/A (2021). PubMed: 33151261
Gao L et al. Thrombopoietin from hepatocytes promotes hematopoietic stem cell regeneration after myeloablation. Elife 10:N/A (2021). PubMed: 34463253
Werder RB et al. Adenine base editing reduces misfolded protein accumulation and toxicity in alpha-1 antitrypsin deficient patient iPSC-hepatocytes. Mol Ther 29:3219-3229 (2021). PubMed: 34217893
Prawira A et al. Ribociclib enhances infigratinib-induced cancer cell differentiation and delays resistance in FGFR-driven hepatocellular carcinoma. Liver Int 41:608-620 (2021). PubMed: 33179425

View all Publications for this product

Customer reviews and Q&As

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Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (Liver)

Permeabilization

Yes - 0.3%TritonX100

Specification

Liver

Blocking step

normal goat serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Oct 09 2018

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Key features and details

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Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (14)

Customer reviews and Q&As

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Immunohistochemistry (Frozen sections) abreview for Anti-HNF-4-alpha antibody [EPR16885-99]

Post-translational
modifications