Overview

  • Product name

  • Description

    Mouse monoclonal to NRF1
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment (GST-tag) corresponding to Human NRF1 aa 201-285.
    Sequence:

    TQAQLRAFIPEMLKYSTGRGKPGWGKESCKPIWWPEDIPWANVRSDVRTE EQKQRVSWTQALRTIVKNCYKQHGREDLLYAFEDQ


    Database link: Q16656

  • Positive control

  • General notes

    This product was changed from ascites to tissue culture supernatant on 15 May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab55744 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 Use at an assay dependent concentration. Predicted molecular weight: 54 kDa.
IHC-P Use at an assay dependent concentration.
IP Use at an assay dependent concentration. PubMed: 24250222

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

  • 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

  • Anti-NRF1 antibody (ab55744) at 1 µg/ml + Human skeletal muscle lysate

    Secondary
    Goat Anti-Mouse IgG (H&L)-HRP Conjugate

    Predicted band size: 54 kDa



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

  • NRF1 antibody (ab55744) used in immunohistochemistry at  µg/ml on formalin fixed and paraffin embedded human kidney.

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

References

This product has been referenced in:

  • Le Berre G  et al. Repression of TERRA Expression by Subtelomeric DNA Methylation Is Dependent on NRF1 Binding. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 31181625) »
  • Ramos J  et al. NRF1 motif sequence-enriched genes involved in ER/PR -ve HER2 +ve breast cancer signaling pathways. Breast Cancer Res Treat 172:469-485 (2018). Read more (PubMed: 30128822) »
See all 16 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (liver)
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Nov 10 2014

Question
Answer

Below please find a new discount code and new expiration date regarding testing of ab55744 in zebrafish. Please let me know if you have any questions.

=====

I am very pleased to hear you would like to accept our offer and test ab55744 in zebrafish. This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for zebrafish and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

Read More
Application
Western blot
Sample
Zebrafish Cell lysate - whole cell (ZEB2J)
Loading amount
20 µg
Specification
ZEB2J
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jul 03 2012

Question
Answer

DISCOUNT CODE: ***
Expiration date: August 19th, 2012

I am very pleased to hear you would like to accept our offer and test ab55744 in zebrafish. This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for zebrafish and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for getting back to me and for providing some further details. It is very much appreciated. It may be worth running human heart or skeletal muscle lysate alongside the samples to see if endogenously expressed NRF1 protein runs differently under the same conditions. Nevertheless, if your customer would prefer to get a replacement vial or a credit note please do let me know. I look forward to hearing from you and hope to solve this problem as soon as possible.

Read More

Question

LOT NUMBER gr27905-1 ORDER NUMBER 901532 DESCRIPTION OF THE PROBLEM Multiple bands and the major bean is not the correct size SAMPLE •Species: human •What’s cell line or tissue: HEK293T cells •Cell extract or Nuclear extract: Cell extract •Purified protein or Recombinant protein: Purified protein PRIMARY ANTIBODY •Species: mouse •Reacts against:Human •At what dilution(s) have you tested this antibody:1/6000 •What dilution buffer was used: 5% milk in TBST buffer •Incubation time: 20 hr •Incubation temperature: 4℃ •What washing steps were done: (TBST 15ml 10 min) x 3 DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED We used the cell extracts which are transfected with full-length NRF-1 and empty vector as positive controls. We transfected three constructs PCMS-NRF-1-EGFP, Flag-NRF-1 and NRF-1-EGFP-N1 into HEK293T cells. The PCMS-NRF-1-EGFP construct contains full-length NRF-1 and EGFP that are driven by different promoters. The Flag-NRF-1 construct transcript full-length NRF-1 and fused with flag tag. The NRF-1-EGFP-N1 construct will be expressed as fusions to the EGFP. ANTIBODY STORAGE CONDITIONS -20℃ SAMPLE PREPARATION •What lysis buffer was used: CSK buffer (10mM HEPES (pH 6.8), 300mM Sucrose, 100mM NaCl, 3mM MgCl2, 1mM EGTA, 0.5% Triton X-100, 0.2mM EDTA, 0.5mM PMSF, 0.5mM DTT, 50mM NaF, 1mM Na3V04, leupeptin, aprotinin) •What protease inhibitors were used: DTT, leupeptin, aprotinin, PMSF, EDTA, EGTA • What loading buffer was used: 20% glycerol, 0.02mg/ml bromophenol blue, 2.5% SDS, 5% 2-mercaptoehtanol, 1*stacking buffer (6.06 g Tris-base/100ml H2O, 0.4% SDS) •Phosphatase inhibitors: NaF, Na3VO4, •Did you heat the samples: temperature and time: 95℃, 5 min AMOUNT OF PROTEIN LOADED 22.5 ug ELECTROPHORESIS/GEL CONDITIONS •Reducing or non reducing gel: reducing gel •Reducing agent: beta-mercaptoethanol •Gel percentage : 10% TRANSFER AND BLOCKING CONDITIONS •Transfer conditions: (Type of membrane, Protein transfer verified): PVDF •Buffer: TBST Buffer •Blocking agent: milk, BSA, serum, what percentage: milk 5% •Incubation time: 1 hr •Incubation temperature: 24℃ SECONDARY ANTIBODY •Species: goat •Reacts against: mouse •At what dilution(s) have you tested this antibody: 30000 •Incubation time: 1 hr at 24℃ •Wash steps: (TBST 15ml 10 min) x 3 •Fluorochrome or enzyme conjugate: HRP •Do you know whether the problems you are experiencing come from the secondary? no HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Reducing primary and secondary antibody concentrations. Reduce the protein loading concentration.

Read More
Answer

Thank you for contacting Abcam Technical Team and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with this product. Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem. 1) Image: a) Could you please specify what size are the non-specific bands? Are they lower or higher than the expected size? b) Could you attach an image (saved as a jpeg file) to your e-mail so that I can have a look at the bands. c) There are at least two isoforms have been described in the literature. Could you please comment on which isoform(s) is expected to be expressed in the samples? 2) Construct: Epitope tagging could interact with the recognition of the epitope(s). a) Where is the location of the tag within the reading frame of the NRF1 protein? b) How many copies of tags have been inserted? c) Has the transfection efficiency been tested? 3) Positive control: Since the target protein is tagged, it would be worth running a good positive control alongside with the samples on the same gel to see if the band pattern is the same or not. As the on-line datasheet indicates, human kidney lysates were used to test this antibody, so this type of lysate or human heart could be used as positive control. Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

Read More
Application
Western blot
Sample
Human Tissue lysate - whole (Muscle, vastus lateralis)
Loading amount
20 µg
Specification
Muscle, vastus lateralis
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Dr. Andrew Murray

Verified customer

Submitted Mar 17 2009

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