RecombinantRabMAb

Recombinant Anti-NFE2 antibody [EPR20463] (ab223845)

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Western blot - Anti-NFE2 antibody [EPR20463] (ab223845)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFE2 antibody [EPR20463] (ab223845)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFE2 antibody [EPR20463] (ab223845)
  • Flow Cytometry - Anti-NFE2 antibody [EPR20463] (ab223845)
  • Anti-NFE2 antibody [EPR20463] (ab223845)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20463] to NFE2
  • Suitable for: WB, IHC-P, Flow Cyt
  • Reacts with: Human

Overview

  • Product name

    Anti-NFE2 antibody [EPR20463]
    See all NFE2 primary antibodies

  • Description

    Rabbit monoclonal [EPR20463] to NFE2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human NFE2 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q16621

  • Positive control

    • WB: K562, HEL and TF-1 whole cell lysates. IHC-P: Human placenta and malignant lymphoma of bone marrow tissue. Flow Cytometry: HEL cells.

  • General notes

     This product was previously labelled as Nuclear Factor Erythroid Derived 2

     

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab223845 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: 41 kDa.
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/400.

Target

  • Function

    Component of the NF-E2 complex essential for regulating erythroid and megakaryocytic maturation and differentiation. Binds to the hypersensitive site 2 (HS2) of the beta-globin control region (LCR). This subunit (NFE2) recognizes the TCAT/C sequence of the AP-1-like core palindrome present in a number of erythroid and megakaryocytic gene promoters. Requires MAFK or other small MAF proteins for binding to the NF-E2 motif. May play a role in all aspects of hemoglobin production from globin and heme synthesis to procurement of iron.

  • Tissue specificity

    Expressed in hematopoietic cells and also in colon and testis.

  • Sequence similarities

    Belongs to the bZIP family. CNC subfamily.
    Contains 1 bZIP domain.

  • Domain

    The PXY motifs are required for binding WW domains. PXY1 is required to promote transactivation of beta-globin and for hyperacetylation of histone H3, but not for binding to the HS2 promoter site.

  • Post-translational
    modifications

    Phosphorylated on serine residues (By similarity). In undifferentiated erythrocytes, phosphorylated by MAPK8 which then leads to ubiquitination and protein degradation.
    Sumoylated. Sumoylation is required for translocation to nuclear bodies PODs, anchoring to the gene loci, and transactivation of the beta-globin gene.
    Ubiquitinated mainly by 'Lys63'-linked ubiquitin. Polyubiquitination with 'Lys63'-linked ubiquitin by ITCH retains NFE2 in the cytoplasm preventing its transactivation activity. In undifferentiated erythrocyte, ubiquitinated after MAPK8-mediatd phosphorylation leading to protein degradation.

  • Cellular localization

    Nucleus > PML body. Cytoplasm. The sumoylated form locates to the nuclear bodies PML oncogenic domains (PODs). Translocated to the cytoplasm through interaction with ITCH.
  • Information by UniProt

  • Database links

    • Alternative names

      • erythroid-derived 2 45 kDa subunit antibody
      • Leucine zipper protein NF-E2 antibody
      • Leucine zipper protein NFE2 antibody
      • NF E2 antibody
      • nfe2 antibody
      • NFE2_HUMAN antibody
      • Nuclear factor (erythroid derived 2), 45kDa antibody
      • nuclear factor (erythroid-derived 2), 45kD antibody
      • Nuclear factor antibody
      • nuclear factor, erythroid 2 antibody
      • Nuclear factor, erythroid derived 2 45 kDa subunit antibody
      • p45 antibody
      • p45 NF-E2 antibody
      • p45 NFE2 antibody
      • Transcription factor NF-E2 45 kDa subunit antibody
      • Transcription factor NFE2 45 kDa subunit antibody
      see all

    Images

    • Western blot - Anti-NFE2 antibody [EPR20463] (ab223845)
      Western blot - Anti-NFE2 antibody [EPR20463] (ab223845)
      All lanes : Anti-NFE2 antibody [EPR20463] (ab223845) at 1/1000 dilution

      Lane 1 : K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
      Lane 2 : HEL (human bone marrow erythroleukemia cell line) whole cell lysate
      Lane 3 : TF-1 (human bone marrow erythroleukemia cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 41 kDa
      Observed band size: 41 kDa


      Exposure time: 3 minutes


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFE2 antibody [EPR20463] (ab223845)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFE2 antibody [EPR20463] (ab223845)

      Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling NFE2 with ab223845 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in neutrophils (arrow) in human placenta (PMID: 8645254) 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) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFE2 antibody [EPR20463] (ab223845)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFE2 antibody [EPR20463] (ab223845)

      Immunohistochemical analysis of paraffin-embedded malignant lymphoma of bone marrow tissue labeling NFE2 with ab223845 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on a subset of cells of malignant lymphoma of bone marrow 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) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Flow Cytometry - Anti-NFE2 antibody [EPR20463] (ab223845)
      Flow Cytometry - Anti-NFE2 antibody [EPR20463] (ab223845)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEL (human bone marrow erythroleukemia) cell line labeling NFE2 with ab223845 at 1/400 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled 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.

    • Anti-NFE2 antibody [EPR20463] (ab223845)
      Anti-NFE2 antibody [EPR20463] (ab223845)

    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

    References (0)

    Publishing research using ab223845? Please let us know so that we can cite the reference in this datasheet.

    ab223845 has not yet been referenced specifically in any publications.

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