Recombinant
RabMAb

Recombinant Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

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Overview

  • Product name

    Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade
    See all NRF1 primary antibodies

  • Description

    Rabbit monoclonal [EPR5554(N)] to NRF1 - ChIP Grade

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ChIP, WB, ICC/IF, Flow Cyt, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human NRF1 aa 350-450 (Cysteine residue). The exact sequence is proprietary.
    Database link: Q16656

  • Positive control

    • WB: MCF-7, HeLa and 293T cell lysates and human fetal heart, mouse heart, mouse brain, rat heart and rat brain tissue lysates. IHC-P: Human gastric adenocarcinoma, human cervical carcinoma and human skeletal muscle tissues. ICC/IF: HeLa and MCF-7 cells. Flow Cyt: 293T cells. IP: 293T cell lysate.

  • 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

Applications

Our Abpromise guarantee covers the use of ab175932 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
ChIP Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Predicted molecular weight: 54 kDa.
ICC/IF 1/50 - 1/100.
Flow Cyt 1/10 - 1/150.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP 1/10 - 1/100.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Target

  • Function

    Transcription factor that activates the expression of the EIF2S1 (EIF2-alpha) gene. Links the transcriptional modulation of key metabolic genes to cellular growth and development. Implicated in the control of nuclear genes required for respiration, heme biosynthesis, and mitochondrial DNA transcription and replication.

  • Tissue specificity

    Ubiquitously expressed with strongest expression in skeletal muscle.

  • Sequence similarities

    Belongs to the NRF1/Ewg family.

  • Post-translational
    modifications

    Phosphorylation enhances DNA binding.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • alpha pal antibody
    • alpha palindromic binding protein antibody
    • Alpha palindromic-binding protein antibody
    • Alpha-pal antibody
    • locus control region factor 1 antibody
    • NFE2 related factor 1 antibody
    • NRF-1 antibody
    • Nrf1 antibody
    • NRF1_HUMAN antibody
    • Nuclear respiratory factor 1 antibody
    see all

Images

  • Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    All lanes : Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/5000 dilution (purified)

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : Mouse brain tissue lysate
    Lane 3 : Rat heart tissue lysate
    Lane 4 : Rat brain tissue lysate

    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: 54 kDa
    Observed band size: 68 kDa
    why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cevical carcinoma tissue labelling NRF1 with purified ab175932 at a dilution of 1/100. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/ Immunofluorescence - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Immunocytochemistry/ Immunofluorescence - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

    Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling NRF1 with purified ab175932 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Flow Cytometry - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Flow Cytometry - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

    Flow Cytometry analysis of 293T cells labelling NRF1 with purified ab175932 at a dilution of 1/150 (red). Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • ChIP - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    ChIP - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Chromatin was prepared from Hela cells according to the Abcam Dual X-ChIP protocol. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab175932 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    Primers and probes are located in the first kb of the transcribed region.
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
  • Immunoprecipitation - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Immunoprecipitation - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

    ab175932 (purified) at a dilution of 1/50 immunoprecipitating NRF1 in 293T whole cell lysate.

    Lane 1 (input): 293T whole cell lysate (10µg)

    Lane 2 (+): ab175932 + 293T whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab175932 in 293T whole cell lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • ChIP - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    ChIP - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Chromatin was prepared from NIH/3T3 treated with MG-132(2uM 16h) cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab175932 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    Primers and probes are located in the first kb of the transcribed region.
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
  • Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/10000 dilution (purified) + HEK293 whole cell lysate at 20 µg

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

    Predicted band size: 54 kDa
    Observed band size: 68 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST
  • Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/10000 dilution (purified) + HeLa whole cell lysate at 20 µg

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

    Predicted band size: 54 kDa
    Observed band size: 68 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST
  • Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Western blot - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    All lanes : Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932) at 1/1000 dilution (unpurified)

    Lane 1 : MCF-7 cell lysate
    Lane 2 : Hela cell lysate
    Lane 3 : Human fetal heart tissue lysate
    Lane 4 : 293T cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 54 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Immunocytochemistry/ Immunofluorescence - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling NRF1 with unpurified ab175932 at a dilution of 1/50.

  • Immunoprecipitation - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Immunoprecipitation - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

    ab175932 (unpurified) at a dilution of 1/10 immunoprecipitating NRF1 in 293T cell lysate.

  • Flow Cytometry - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Flow Cytometry - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

    Flow cytometric analysis of permeabilized 293T cells labeling NRF1 with unpurified ab175932 at a dilution of 1/10 (red) compared to a negative control (rabbit IgG, green).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue labeling NRF1 with unpurified ab175932 at a dilution of 1/50.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody [EPR5554(N)] - ChIP Grade (ab175932)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric adenocarcinoma tissue labeling NRF1 with unpurified ab175932 at a dilution of 1/50.

References

This product has been referenced in:

  • Niu N  et al. Effects of nuclear respiratory factor-1 on apoptosis and mitochondrial dysfunction induced by cobalt chloride in H9C2 cells. Mol Med Rep 19:2153-2163 (2019). Read more (PubMed: 30628711) »
  • Geng J  et al. TIGAR regulates mitochondrial functions through SIRT1-PGC1a pathway and translocation of TIGAR into mitochondria in skeletal muscle. FASEB J 33:6082-6098 (2019). Read more (PubMed: 30726106) »
See all 17 Publications for this product

Publishing research using ab175932? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-NRF1 antibody [EPR5554(N)]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (MCF-7)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
MCF-7
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Aug 09 2016

Western blot abreview for Anti-NRF1 antibody [EPR5554(N)]

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (12% tris-glycine gel)
Sample
Human Tissue lysate - whole (heart)
Specification
heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More

Abcam user community

Verified customer

Submitted Oct 10 2014

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