Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
- Datasheet
- References (4)
- Protocols
Overview
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Product nameAnti-HNF-4-alpha antibody [EPR16885] - ChIP Grade
See all HNF-4-alpha primary antibodies -
DescriptionRabbit monoclonal [EPR16885] to HNF-4-alpha - ChIP Grade
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Host speciesRabbit
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Tested applicationsSuitable for: ChIP, IHC-P, WB, IPmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Recombinant fragment within Mouse HNF-4-alpha aa 1-200. The exact sequence is proprietary.
Database link: P49698 -
Positive control
- WB: HepG2 and SW480 whole cell lysates; Human fetal liver, colon and fetal kidney lysates; mouse and rat liver lysates. IHC-P: Human liver, Human colon, mouse liver and rat colon tissues. IP: HepG2 whole cell extract. ChIP: HepG2 whole cell extract.
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General 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 rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferPreservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEPR16885
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IsotypeIgG
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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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ChIP Related Products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab181604 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 |
|---|---|---|
| ChIP | Use 2 µg for 25 µg of chromatin. | |
| IHC-P | 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
| WB | 1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa). | |
| IP | 1/70. |
Target
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FunctionTranscriptionally 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.
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Involvement in diseaseDefects 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.
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Sequence similaritiesBelongs 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 localizationNucleus.
- Information by UniProt
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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
see all -
Alternative names
- FLJ39654 antibody
- FRTS4 antibody
- Hepatic nuclear factor 4 alpha antibody
see all
Images
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![ChIP - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-245745-Untitled.jpg)
ChIP - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)Chromatin was prepared from HepG2 (Human liver hepatocellular carcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab181604 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
ABCC6_NC1 is negative control
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![Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-271945-Publish-image-HNF4a.jpg)
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)All lanes : Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/10000 dilution
Lane 1 : HepG2 whole cell lysate
Lane 2 : HEK293 whole cell lysate
Lane 3 : Mouse liver lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution
Predicted band size: 53 kDaThis blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour before being incubated with ab181604 (rabbit monoclonal [EPR16885] to HNF-4-alpha; dilution 1:10000) and loading control ab18058 (mouse monoclonal [SPM227] to Vinculin; dilution 1:10000) at 4°C overnight. Antibody binding was detected with ab216773 (Goat anti-Rabbit IgG H&L (IRDye® 800CW); green; dilution 1:20000) and ab216776 (Goat anti-Mouse IgG H&L (IRDye® 680RD), red; dilution 1:20000) for 1 hour at room temperature before imaging.
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![Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242418-wb-1.jpg)
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) 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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242417-wb-2.jpg)
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) 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 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242416-wb-3.jpg)
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution + SW480 (Human colon adenocarcinoma cell line) 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: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242415-wb-4.jpg)
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)All lanes : Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : Human colon lysate
Lane 3 : 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 minuteThis antibody can recognize 6 isoforms in human. The predicted MW are 53KDa, 52 KDa, 47KDa, 56 KDa, 49 KDa and 44 KDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242414-wb-5.jpg)
Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)All lanes : Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution
Lane 1 : Mouse liver lysate
Lane 2 : Rat liver lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 53 kDa
Exposure time: 15 secondsThis antibody can recognize 2 isoforms in mouse and rat. The predicted MW are 53KDa and 52 KDa.
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242413-181604IHCa.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Human liver 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242412-181604IHCb.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on epithelial cells of Human colon 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242411-181604IHCc.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HNF-4-alpha with ab181604 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 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242410-181604IHCd.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on epithelial cells of rat colon 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.
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![Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)](https://www.abcam.com/ps/products/181/ab181604/Images/ab181604-242409-IP.jpg)
Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)HNF-4 alpha was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell extract with ab181604 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab181604 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: HepG2 whole cell extract 10 µg (Input).
Lane 2: ab181604 IP in HepG2 whole cell extract.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181604 in HepG2 whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Protocols
Datasheets and documents
References
This product has been referenced in:
- Zhang N et al. Elevated hepatic expression of H19 long noncoding RNA contributes to diabetic hyperglycemia. JCI Insight 3:N/A (2018). Read more (PubMed: 29769440) »
- Jia Y et al. Dapagliflozin Aggravates Renal Injury via Promoting Gluconeogenesis in db/db Mice. Cell Physiol Biochem 45:1747-1758 (2018). Read more (PubMed: 29495021) »
