Overview

  • Product name
    Anti-NELFe antibody [EPR11600]
    See all NELFe primary antibodies
  • Description
    Rabbit monoclonal [EPR11600] to NELFe
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IP, Flow Cytmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Human NELFe.

  • Positive control
    • WB: HeLa, T47-D, HepG2 or 293T lysate, HepG2 cells. Mouse and Rat brain lysates. ICC/IF: HeLa cells. FC: HeLa cells. IP: HeLa cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

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 long term.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR11600
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab170104 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 - 1/5000. Predicted molecular weight: 44 kDa.
ICC/IF 1/500.

For unpurified use at 1/100 - 1/250.

IP 1/10 - 1/100.
Flow Cyt 1/300.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function
      Essential component of the NELF complex, a complex that negatively regulates the elongation of transcription by RNA polymerase II. The NELF complex, which acts via an association with the DSIF complex and causes transcriptional pausing, is counteracted by the P-TEFb kinase complex.
    • Tissue specificity
      Widely expressed. Expressed in heart, brain, lung, placenta, liver, skeletal muscle, kidney and pancreas.
    • Sequence similarities
      Belongs to the RRM NELF-E family.
      Contains 1 RRM (RNA recognition motif) domain.
    • Domain
      The RRM domain interacts with RNA, and is essential for NELF complex function. It is however not required for the NELF complex formation.
    • Post-translational
      modifications
      Sumoylated.
    • Cellular localization
      Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • Major histocompatibility complex gene RD antibody
      • Negative elongation factor E antibody
      • Negative elongation factor polypeptide E antibody
      • NELF e antibody
      • NELF-E antibody
      • Nelfe antibody
      • NELFE_HUMAN antibody
      • Nuclear protein antibody
      • RD antibody
      • RD protein antibody
      • RD RNA binding protein antibody
      • RDP antibody
      • RNA-binding protein RD antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling NELFe with Purified ab170104 at 1:500 dilution (0.5 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • ab170104 (purified) at 1:20 dilution (2µg) immunoprecipitating NELFe in HeLa whole cell lysate.
      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab170104 & HeLa whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab170104 in HeLa whole cell lysate
      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling NELFe with Purified ab170104 at 1:300 dilution (1 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • All lanes : Anti-NELFe antibody [EPR11600] (ab170104) at 1/5000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : Mouse brain lysates
      Lane 3 : Rat brain lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 44 kDa
      Observed band size: 44 kDa

    • All lanes : Anti-NELFe antibody [EPR11600] (ab170104) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : T47-D cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : 293T cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit
      at 1/2000 dilution

      Predicted band size: 44 kDa



      This image was generated using the unpurified version of the product. 

    • Immunofluorescence analysis of HepG2 cells labeling NELFe with ab170104 at a 1/100 dilution.

      This image was generated using the unpurified version of the product. 

    • Western blot analysis on immunoprecipitation pellet from HeLa cell lysate (lane 1) or 1X PBS (negative control) (lane 2) using ab170104, and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

      This image was generated using the unpurified version of the product. 

    References

    This product has been referenced in:
    • Zhao Z  et al. CDK9 and SPT5 proteins are specifically required for expression of herpes simplex virus 1 replication-dependent late genes. J Biol Chem 292:15489-15500 (2017). Read more (PubMed: 28743741) »
    • Awwad SW  et al. NELF-E is recruited to DNA double-strand break sites to promote transcriptional repression and repair. EMBO Rep 18:745-764 (2017). Read more (PubMed: 28336775) »
    See all 2 Publications for this product

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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