Recombinant Anti-NUR77 antibody [EPR23982-85] (ab283264)
![Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)](https://www.abcam.com/ps/products/283/ab283264/Images/ab283264-477270-anti-nur77-antibody-epr23982-85-western-blot-wild-type-hela-treated-mg132-10-um-6-h-nr4a1-knockout-hela-treated-mg.jpg "Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23982-85] to NUR77
- Suitable for: WB, Flow Cyt (Intra), IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-NUR77 antibody [EPR23982-85]
See all NUR77 primary antibodies -
Description
Rabbit monoclonal [EPR23982-85] to NUR77 -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt (Intra), IPmore details
Unsuitable for: ICC/IF or IHC-P -
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: Untreated Jurkat whole cell lysate; Jurkat treated 40nM TPA and 2µM calcium ionophore A23187 for 4 hours whole cell lysate; Jurkat treated 40nM TPA, 2µM calcium ionophore A23187 and 10µM MG-132 for 4 hours whole cell lysate; Untreated HeLa whole cell lysate; HeLa treated with 10µM MG-132 for 6h whole cell lysate. Flow Cyt(intra): Jurkat cells. IP: HeLa treated with 10µM MG-132 for 6h whole cell lysate.
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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.
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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23982-85 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab283264 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 |
1/1000. Predicted molecular weight: 64 kDa.
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| Flow Cyt (Intra) |
1/500.
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| IP |
1/30.
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| Notes |
|---|
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WB
1/1000. Predicted molecular weight: 64 kDa. |
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Flow Cyt (Intra)
1/500. |
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IP
1/30. |
Target
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Function
Orphan nuclear receptor. May act concomitantly with NURR1 in regulating the expression of delayed-early genes during liver regeneration. Binds the NGFI-B response element (NBRE) 5'-AAAAGGTCA-3' (By similarity). May inhibit NF-kappa-B transactivation of IL2. -
Tissue specificity
Fetal muscle and adult liver, brain and thyroid. -
Sequence similarities
Belongs to the nuclear hormone receptor family. NR4 subfamily.
Contains 1 nuclear receptor DNA-binding domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 3164 Human
- Omim: 139139 Human
- SwissProt: P22736 Human
- Unigene: 524430 Human
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Alternative names
- Early response protein NAK1 antibody
- GFRP 1 antibody
- GFRP antibody
see all
Images
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Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)All lanes : Anti-NUR77 antibody [EPR23982-85] (ab283264) at 1/1000 dilution
Lane 1 : Wild type HeLa treated MG132 (10 uM, 6 h) cell lysate
Lane 2 : NR4A1 knockout HeLa treated MG132 (10 uM, 6 h) cell lysate
Lane 3 : Wild type HeLa vehicle control MG132 (0 uM, 6 h) cell lysate
Lane 4 : NR4A1 knockout HeLa vehicle control MG132 (0 uM, 6 h) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 64 kDa
Observed band size: 62-70 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-NUR77 antibody [EPR23982-85] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab283264 was shown to bind specifically to NUR77. A band was observed at 62-70 kDa in treated wild-type HeLa cell lysates with no signal observed at this size in NR4A1 knockout cell line ab264021 (knockout cell lysate ab264509). To generate this image, wild-type and NR4A1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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![Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)](https://www.abcam.com/ps/products/283/ab283264/Images/ab283264-469334-anti-nur77-antibody-epr2398285-western-blot-t.jpg)
Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)All lanes : Anti-NUR77 antibody [EPR23982-85] (ab283264) at 1/1000 dilution
Lane 1 : Untreated Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : Jurkat treated 40nM TPA and 2uM calcium ionophore A23187 for 4 hours whole cell lysate
Lane 3 : Jurkat treated 40nM TPA, 2uM calcium ionophore A23187 and 10uM MG-132 for 4 hours whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 64 kDa
Observed band size: 32,38,60-80 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID: 30224829, 28111880, 29669342).
Exposure time: 15 seconds.
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![Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)](https://www.abcam.com/ps/products/283/ab283264/Images/ab283264-469335-anti-nur77-antibody-epr2398285-western-blot-cervix.jpg)
Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)All lanes : Anti-NUR77 antibody [EPR23982-85] (ab283264) at 1/1000 dilution
Lane 1 : Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa treated with 10uM MG-132 for 6h whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 64 kDa
Observed band size: 32,38,60-80 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID: 30224829, 28111880, 29669342).
Exposure time: 15 seconds.
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![Flow Cytometry - Anti-NUR77 antibody [EPR23982-85] (ab283264)](https://www.abcam.com/ps/products/283/ab283264/Images/ab283264-1-anti-nur77-antibody-epr2398285-flow-cytometry-jurkat.jpg)
Flow Cytometry - Anti-NUR77 antibody [EPR23982-85] (ab283264)Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte) cells treated with 40nM TPA and 2 µM calcium ionophore for 4 hours (Red) or untreated control (Green) cells labelling NUR77 with ab283264 at 1/500 dilution (0.1 µg / Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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![Immunoprecipitation - Anti-NUR77 antibody [EPR23982-85] (ab283264)](https://www.abcam.com/ps/products/283/ab283264/Images/ab283264-2-anti-nur77-antibody-epr2398285-immunoprecipitation-hela.jpg)
Immunoprecipitation - Anti-NUR77 antibody [EPR23982-85] (ab283264)NUR77 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) cells treated with 10µM MG-132 for 6h whole cell lysate with ab283264 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283264 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: HeLa cells treated with 10µM MG-132 for 6h whole cell lysate 10 µg
Lane 2: ab283264 IP in HeLa cells treated with 10µM MG-132 for 6h whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283264 in HeLa treated with 10µM MG-132 for 6h whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds.
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![Anti-NUR77 antibody [EPR23982-85] (ab283264)](https://www.abcam.com/ps/products/283/ab283264/Images/ab283264-3-Benefits-of-Recombinant-antibodies.jpg)
Anti-NUR77 antibody [EPR23982-85] (ab283264)
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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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab283264 has not yet been referenced specifically in any publications.