Recombinant Anti-HNF-4-alpha antibody [EPR3648] (ab92378)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3648] to HNF-4-alpha
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra), ChIC/CUT&RUN-seq, ChIP-sequencing
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-HNF-4-alpha antibody [EPR3648]
See all HNF-4-alpha primary antibodies -
Description
Rabbit monoclonal [EPR3648] to HNF-4-alpha -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra), ChIC/CUT&RUN-seq, ChIP-sequencingmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HepG2, A549 and SW480 cell lysates. IHC-P: Human colon and kidney tissues. ICC/IF: HepG2 cells. Flow Cyt (intra): HepG2 cells. ChIP-seq: HepG2 cells. ChIC/CUT&RUN-Seq: HepG2 cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3648 -
Isotype
IgG -
Research areas
- Epigenetics and Nuclear Signaling
- Transcription
- Polymerase associated factors
- Pol II Transcription
- Other
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipid metabolism
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Cholesterol Metabolism
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipases
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92378 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 |
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WB | (1) |
1/2000. Predicted molecular weight: 53 kDa.
For unpurified use at 1/1000 - 1/10000. |
IHC-P | (1) |
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
ICC/IF | (2) |
1/100 - 1/250.
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Flow Cyt (Intra) |
1/70.
For unpurified use at 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5 µg |
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ChIP-sequencing |
Use at an assay dependent concentration.
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Notes |
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WB
1/2000. Predicted molecular weight: 53 kDa. For unpurified use at 1/1000 - 1/10000. |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
ICC/IF
1/100 - 1/250. |
Flow Cyt (Intra)
1/70. For unpurified use at 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5 µg |
ChIP-sequencing
Use at an assay dependent concentration. |
Target
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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
modificationsPhosphorylated 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. - Information by UniProt
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Database links
- Entrez Gene: 3172 Human
- Omim: 600281 Human
- SwissProt: P41235 Human
- Unigene: 116462 Human
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Alternative names
- FLJ39654 antibody
- FRTS4 antibody
- Hepatic nuclear factor 4 alpha antibody
see all
Images
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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 ab92378 [EPR3648]. 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.
The ChIP data was conducted on chromatin prepared from HepG2 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HepG2 cells and 8 µg of ab92378. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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Anti-HNF-4-alpha antibody [EPR3648] (ab92378) at 1/2000 dilution (purified) + SW480 cell lysate at 20 µg
Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDaBlocking and dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling HNF-4-alpha with purified ab92378 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling HNF-4 with purified ab92378 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/500) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
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Anti-HNF-4-alpha antibody [EPR3648] (ab92378) at 1/5000 dilution (purified) + HepG2 cell lysate at 10 µg
Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDaBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-HNF-4-alpha antibody [EPR3648] (ab92378) at 1/1000 dilution (unpurified)
Lane 1 : HepG2 cell lysate
Lane 2 : A549 cell lysate
Lane 3 : SW480 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 53 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue (A) and human kidney tissue (B) labelling HNF-4-aplha with unpurified ab92378 at a 1/100 dilution. Detection: DAB staining. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
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Immunocytochemistry/Immunfluorescence analysis of HepG2 cells labelling HNF-4-alpha with unpurified ab92378 at a 1/100 dilution.
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Intracellular Flow Cytometry analysis of HepG2 cells labelling HNF-4 with purified ab92378 at 1/70 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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Overlay histogram showing HepG2 cells stained with unpurifiedab92378 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (unpurified ab92378, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Chromatin was prepared from HepG2 (Human liver hepatocellular carcinoma cell line) cells. ChIP was performed with 10^7 HepG2 cells and 8 µg of ab92378 [EPR3648]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (14)
ab92378 has been referenced in 14 publications.
- Wang N et al. HDAC6/HNF4a loop mediated by miR-1 promotes bile acids-induced gastric intestinal metaplasia. Gastric Cancer 24:103-116 (2021). PubMed: 32705446
- Borys BS et al. Overcoming bioprocess bottlenecks in the large-scale expansion of high-quality hiPSC aggregates in vertical-wheel stirred suspension bioreactors. Stem Cell Res Ther 12:55 (2021). PubMed: 33436078
- Huang KW et al. Liver Activation of Hepatocellular Nuclear Factor-4a by Small Activating RNA Rescues Dyslipidemia and Improves Metabolic Profile. Mol Ther Nucleic Acids 19:361-370 (2020). PubMed: 31877412
- Famulari ES et al. Human liver stem cells express UGT1A1 and improve phenotype of immunocompromised Crigler Najjar syndrome type I mice. Sci Rep 10:887 (2020). PubMed: 31965023
- Rohani L et al. Stirred suspension bioreactors maintain naïve pluripotency of human pluripotent stem cells. Commun Biol 3:492 (2020). PubMed: 32895477