RecombinantRabMAb

Recombinant Anti-NELFe antibody [EPR11600] (ab170104)

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Immunocytochemistry/ Immunofluorescence - Anti-NELFe antibody [EPR11600] (ab170104)
  • Immunoprecipitation - Anti-NELFe antibody [EPR11600] (ab170104)
  • Flow Cytometry (Intracellular) - Anti-NELFe antibody [EPR11600] (ab170104)
  • Western blot - Anti-NELFe antibody [EPR11600] (ab170104)
  • Western blot - Anti-NELFe antibody [EPR11600] (ab170104)
  • Immunocytochemistry/ Immunofluorescence - Anti-NELFe antibody [EPR11600] (ab170104)
  • Immunoprecipitation - Anti-NELFe antibody [EPR11600] (ab170104)
  • Anti-NELFe antibody [EPR11600] (ab170104)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR11600] to NELFe
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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: Flow Cyt (Intra), WB, ICC/IF, IPmore 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. Flow Cyt (intra): HeLa cells. IP: HeLa cells.

  • 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.

    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.

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 (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR11600

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/300.
WB (1)
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.
Notes
Flow Cyt (Intra)
1/300.
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.
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

    see all

  • 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 - Anti-NELFe antibody [EPR11600] (ab170104)
    Immunocytochemistry/ Immunofluorescence - Anti-NELFe antibody [EPR11600] (ab170104)

    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.

  • Immunoprecipitation - Anti-NELFe antibody [EPR11600] (ab170104)
    Immunoprecipitation - Anti-NELFe antibody [EPR11600] (ab170104)

    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 (Intracellular) - Anti-NELFe antibody [EPR11600] (ab170104)
    Flow Cytometry (Intracellular) - Anti-NELFe antibody [EPR11600] (ab170104)

    Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling NELFe with Purified ab170104 at 1/300 dilution (1µg/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). 

     

     

  • Western blot - Anti-NELFe antibody [EPR11600] (ab170104)
    Western blot - Anti-NELFe antibody [EPR11600] (ab170104)
    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

  • Western blot - Anti-NELFe antibody [EPR11600] (ab170104)
    Western blot - Anti-NELFe antibody [EPR11600] (ab170104)
    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. 

  • Immunocytochemistry/ Immunofluorescence - Anti-NELFe antibody [EPR11600] (ab170104)
    Immunocytochemistry/ Immunofluorescence - Anti-NELFe antibody [EPR11600] (ab170104)

    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. 

  • Immunoprecipitation - Anti-NELFe antibody [EPR11600] (ab170104)
    Immunoprecipitation - Anti-NELFe antibody [EPR11600] (ab170104)

    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. 

  • Anti-NELFe antibody [EPR11600] (ab170104)
    Anti-NELFe antibody [EPR11600] (ab170104)

References (9)

Publishing research using ab170104? Please let us know so that we can cite the reference in this datasheet.

ab170104 has been referenced in 9 publications.

  • Takahashi H  et al. The role of Mediator and Little Elongation Complex in transcription termination. Nat Commun 11:1063 (2020). PubMed: 32102997
  • Aoi Y  et al. NELF Regulates a Promoter-Proximal Step Distinct from RNA Pol II Pause-Release. Mol Cell 78:261-274.e5 (2020). PubMed: 32155413
  • Etchegaray JP  et al. The Histone Deacetylase SIRT6 Restrains Transcription Elongation via Promoter-Proximal Pausing. Mol Cell 75:683-699.e7 (2019). PubMed: 31399344
  • Han L  et al. NELFE promoted pancreatic cancer metastasis and the epithelial-to-mesenchymal transition by decreasing the stabilization of NDRG2 mRNA. Int J Oncol 55:1313-1323 (2019). PubMed: 31638184
  • Zhu J  et al. Regulation of Pol II Pausing Is Involved in Daily Gene Transcription in the Mouse Liver. J Biol Rhythms 33:350-362 (2018). PubMed: 29845885
  • 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). 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). PubMed: 28336775
  • Dang H  et al. Oncogenic Activation of the RNA Binding Protein NELFE and MYC Signaling in Hepatocellular Carcinoma. Cancer Cell 32:101-114.e8 (2017). PubMed: 28697339
  • Rother S  et al. NF-?B-repressing factor phosphorylation regulates transcription elongation via its interactions with 5'?3' exoribonuclease 2 and negative elongation factor. FASEB J 30:174-85 (2016). PubMed: 26340924

Customer reviews and Q&As

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Western blot abreview for Anti-NELFe antibody [EPR11600]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (hepatocellular carcinoma cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris Gel)
Loading amount
2 µg
Specification
hepatocellular carcinoma cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 02 2019

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