Recombinant Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)
![ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)](https://www.abcam.com/ps/products/251/ab251275/Images/ab251275-517368-anti-hnf-4-alpha-antibody-epr16786-n-terminal-chic-cut-and-run-sequencing.jpg "ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16786] to HNF-4-alpha - BSA and Azide free
- Suitable for: ICC/IF, WB, IHC-P, ChIC/CUT&RUN-seq, ChIP-sequencing
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free
See all HNF-4-alpha primary antibodies -
Description
Rabbit monoclonal [EPR16786] to HNF-4-alpha - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IHC-P, ChIC/CUT&RUN-seq, ChIP-sequencingmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Mouse and rat liver lysates. Human fetal liver and Human stomach lysates. HepG2 whole cell lysate. IHC-P: Human hepatocellular carcinoma tissue. ICC/IF: HepG2 cells. ChIP-seq: HepG2 cells. ChIC/CUT&RUN-Seq: HepG2 cells.
-
General notes
ab251275 is the carrier-free version of ab199431.
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
-
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
EPR16786 -
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
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab251275 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 |
|---|---|---|
| ICC/IF |
Use at an assay dependent concentration.
|
|
| WB |
Use at an assay dependent concentration. Detects a band of approximately 47, 53 kDa (predicted molecular weight: 53 kDa).
|
|
| IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
| ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
|
|
| ChIP-sequencing |
Use at an assay dependent concentration.
|
| Notes |
|---|
|
ICC/IF
Use at an assay dependent concentration. |
|
WB
Use at an assay dependent concentration. Detects a band of approximately 47, 53 kDa (predicted molecular weight: 53 kDa). |
|
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
|
ChIP-sequencing
Use at an assay dependent concentration. |
Target
-
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
-
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
-
ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)
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 ab199431 [EPR16786]. 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. ChIP was performed with 10^7 HepG2 cells and 8 µg of ab199431 [EPR16786]. 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.
This data was developed using ab199431, the same antibody clone in a different buffer formulation.
-
![Western blot - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)](https://www.abcam.com/ps/products/251/ab251275/Images/ab251275-408158-anti-hnf-4-alpha-antibody-epr16786-bsa-and-azide-free-western-blot.jpg)
Western blot - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)All lanes : Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431) at 1/10000 dilution
Lane 1 : Mouse liver tissue lysate
Lane 2 : Rat liver tissue lysate
Lane 3 : Human fetal liver tissue lysate
Lane 4 : Human stomach tissue 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: 47, 53 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesThis data was developed using ab199431, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
This antibody can recognize 3 isoforms. The predicted MW are 53KDa, 52 KDa and 47KDa, respectively.
-
![Western blot - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)](https://www.abcam.com/ps/products/251/ab251275/Images/ab251275-405828-anti-hnf-4-alpha-antibody-epr16786-bsa-and-azide-free-western-blot.jpg)
Western blot - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431) 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: 47, 53 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteThis data was developed using ab199431, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
This antibody can recognize 3 isoforms. The predicted MW are 53KDa, 52 KDa and 47KDa, respectively.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)](https://www.abcam.com/ps/products/251/ab251275/Images/ab251275-2-anti-hnf-4-alpha-antibody-epr16786-bsa-and-azide-free-immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)This data was developed using ab199431, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling HNF-4-alpha with ab199431 at 1/400 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on Human hepatocellular carcinoma tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051). Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. -
![Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)](https://www.abcam.com/ps/products/251/ab251275/Images/ab251275-1-anti-hnf-4-alpha-antibody-epr16786-bsa-and-azide-free-immunocytochemistry.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)This data was developed using ab199431, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling HNF-4-alpha with ab199431 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on HepG2 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:-
-ve control 1 - ab199431 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
![ChIP-sequencing - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)](https://www.abcam.com/ps/products/251/ab251275/Images/ab251275-493624-anti-hnf-4-alpha-antibody-epr16786-n-terminal-chip-sequencing-human.jpg)
ChIP-sequencing - Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)Chromatin was prepared from HepG2 (Human liver hepatocellular carcinoma cell line) cells. ChIP was performed with 10^7 HepG2 cells and 8 µg of ab199431 [EPR16786]. 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.
This data was developed using ab199431, the same antibody clone in a different buffer formulation. -
![Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)](https://www.abcam.com/ps/products/251/ab251275/Images/ab251275-5-benefits-of-recombinant-antibodies.png)
Anti-HNF-4-alpha antibody [EPR16786] - BSA and Azide free (ab251275)
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
-
Datasheet download
Certificate of Compliance
References (0)
ab251275 has not yet been referenced specifically in any publications.
