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), 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), 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.
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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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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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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. |
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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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) - Anti-HNF-4-alpha antibody [EPR3648] (ab92378)
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) - Anti-HNF-4-alpha antibody [EPR3648] (ab92378)
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