Recombinant Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
![ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)](https://www.abcam.com/ps/products/226/ab226487/Images/ab226487-517355-anti-hnf-4-alpha-antibody-epr19265-130-chip-grade-chic-cut-and-run-sequencing.jpg "ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19265-130] to HNF-4-alpha - ChIP Grade – BSA and Azide free
- Suitable for: WB, ICC/IF, ChIP, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seq
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free
See all HNF-4-alpha primary antibodies -
Description
Rabbit monoclonal [EPR19265-130] to HNF-4-alpha - ChIP Grade – BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, ChIP, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seqmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human colon and fetal liver tissue lysates; HepG2, Caco-2 and SW480 whole cell lysates. ICC/IF: HepG2 and SW480 cells. Flow Cyt (intra): HepG2 cells. ChIP: HepG2 cells. IP: HepG2 whole cell lysate. ChIC/CUT&RUN-Seq: HepG2 cells.
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General notes
ab226487 is the carrier-free version of ab200142.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19265-130 -
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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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab226487 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 |
|---|---|---|
| WB |
Use at an assay dependent concentration. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
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| ICC/IF |
Use at an assay dependent concentration.
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| ChIP |
Use at an assay dependent concentration.
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| IP |
Use at an assay dependent concentration.
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| Flow Cyt (Intra) |
Use at an assay dependent concentration.
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| ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
|
| Notes |
|---|
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WB
Use at an assay dependent concentration. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa). |
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ICC/IF
Use at an assay dependent concentration. |
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ChIP
Use at an assay dependent concentration. |
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IP
Use at an assay dependent concentration. |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
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ChIC/CUT&RUN-seq
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 sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2 x 10^5 HepG2 (Human liver hepatocellular carcinoma cell line) cells and 5 µg of ab200142 [EPR19265-130]. 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200142).
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![ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)](https://www.abcam.com/ps/products/226/ab226487/Images/ab226487-324657-anti-hnf-4-alpha-antibody-epr19265-130-chip-grade-chip.jpg)
ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)Chromatin was prepared from HepG2 (human hepatocellular carcinoma epithelial cell) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 5 µg of ab200142 (blue), and 20 µl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach).
ChIP was performed according to the literature (PMID: 18850323).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200142).
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![Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)](https://www.abcam.com/ps/products/226/ab226487/Images/ab226487-324659-anti-hnf-4-alpha-antibody-epr19265-130-chip-grade-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SW480 (human colorectal adenocarcinoma cell line) cells labeling HNF-4-alpha with ab200142 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing nuclear staining on SW480 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200142).
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![Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)](https://www.abcam.com/ps/products/226/ab226487/Images/ab226487-324658-anti-hnf-4-alpha-antibody-epr19265-130-chip-grade-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cell line labeling HNF-4-alpha with ab200142 at 1/600 (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200142).
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![Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)](https://www.abcam.com/ps/products/226/ab226487/Images/ab226487-324656-anti-hnf-4-alpha-antibody-epr19265-130-chip-grade-immunoprecipitation.jpg)
Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)HNF-4-alpha was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab200142 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab200142 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) 10 μg (Input).
Lane 2: ab200142 IP in HepG2 whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200142 in HepG2 whole cell lysate (-).
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200142).
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![Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)](https://www.abcam.com/ps/products/226/ab226487/Images/ab226487-287578-anti-hnf-4-alpha-antibody-epr19265-130-chip-grade-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells labeling HNF-4-alpha with ab200142 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing nuclear staining on HepG2 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200142).
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![Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)](https://www.abcam.com/ps/products/226/ab226487/Images/ab226487-6-benefits-of-recombinant-antibodies.png)
Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade - BSA and Azide free (ab226487)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab226487 has not yet been referenced specifically in any publications.
