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  1. Link

    nur77-antibody-epr23982-85-ab283264.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-NUR77 antibody [EPR23982-85] (ab283264)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)
  • Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)
  • Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)
  • Flow Cytometry - Anti-NUR77 antibody [EPR23982-85] (ab283264)
  • Immunoprecipitation - Anti-NUR77 antibody [EPR23982-85] (ab283264)
  • 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

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Overview

  • 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.

  • 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.
  • 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
    For more information see here.

    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 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

    • Cell Biology
    • Apoptosis
    • Nucleus
    • Other
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Co-activators/co-repressors
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Orphan Nuclear Receptors
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Other
    • Cancer
    • Cell Death
    • Apoptosis
    • Other
    • Cancer
    • Cell Death
    • Apoptosis
    • Nucleus
    • Other

Associated products

  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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.
Flow Cyt (Intra)
1/500.
IP
1/30.
Notes
WB
1/1000. Predicted molecular weight: 64 kDa.
Flow Cyt (Intra)
1/500.
IP
1/30.
Application notes
Is unsuitable for ICC/IF or IHC-P.

Target

  • 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.
  • Target information above from: UniProt accession P22736 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3164 Human
    • Omim: 139139 Human
    • SwissProt: P22736 Human
    • Unigene: 524430 Human
    • Alternative names

      • Early response protein NAK1 antibody
      • GFRP 1 antibody
      • GFRP antibody
      • GFRP1 antibody
      • Growth factor inducible nuclear protein N10 antibody
      • Growth Factor Inducible Nuclear Protein NP10 antibody
      • Growth Factor Response Protein 1 antibody
      • Hbr1 antibody
      • HMR antibody
      • Hormone Receptor antibody
      • MGC9485 antibody
      • N10 antibody
      • N10 nuclear protein antibody
      • NAK 1 antibody
      • NAK1 antibody
      • Nerve growth factor IB nuclear receptor variant 1 antibody
      • NGFIB antibody
      • NP 10 antibody
      • NP10 antibody
      • NR4A1 antibody
      • NR4A1_HUMAN antibody
      • Nuclear hormone receptor NUR/77 antibody
      • Nuclear Hormone Receptor TR3 antibody
      • Nuclear receptor subfamily 4 group A member 1 antibody
      • NUR77 antibody
      • NUR77, mouse, homolog of antibody
      • Orphan nuclear receptor HMR antibody
      • Orphan nuclear receptor NR4A1 antibody
      • Orphan nuclear receptor TR3 antibody
      • Orphan receptor tr3 antibody
      • Receptor NGFIB antibody
      • ST 59 antibody
      • ST-59 antibody
      • ST59 antibody
      • Steroid receptor TR3 antibody
      • Testicular receptor 3 antibody
      • TR 3 antibody
      • TR3 antibody
      • TR3 orphan receptor antibody
      see all

    Images

    • Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)
      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.

    • Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)
      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.

    • Western blot - Anti-NUR77 antibody [EPR23982-85] (ab283264)
      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.

    • Flow Cytometry - Anti-NUR77 antibody [EPR23982-85] (ab283264)
      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.

    • Immunoprecipitation - Anti-NUR77 antibody [EPR23982-85] (ab283264)
      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.

    • Anti-NUR77 antibody [EPR23982-85] (ab283264)
      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.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab283264? Please let us know so that we can cite the reference in this datasheet.

    ab283264 has not yet been referenced specifically in any publications.

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