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    nrf1-antibody-ab34682.pdf

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Epigenetics and Nuclear Signaling Transcription Polymerase associated factors Pol II Transcription Other
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Anti-NRF1 antibody (ab34682)

  • Datasheet
  • SDS
Reviews (3)Q&A (7)References (32)

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Western blot - Anti-NRF1 antibody (ab34682)
  • Western blot - Anti-NRF1 antibody (ab34682)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody (ab34682)
  • Immunocytochemistry/ Immunofluorescence - Anti-NRF1 antibody (ab34682)

Key features and details

  • Rabbit polyclonal to NRF1
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-NRF1 antibody
    See all NRF1 primary antibodies
  • Description

    Rabbit polyclonal to NRF1
  • Host species

    Rabbit
  • Specificity

    ab34682 recognises NRF1 (also known as nuclear respiratory factor 1, alpha palindromic binding protein and alpha-pal).
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Recombinant full length protein, corresponding to amino acids 1-534 of Mouse NRF1

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Polymerase associated factors
    • Pol II Transcription
    • Other
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial Biogenesis
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Nucleotide metabolism
    • Molecular processes
    • Mitochondrial transcription

Associated products

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
    • HeLa whole cell lysate (ab29545)
  • Recombinant Protein

    • Recombinant Human NRF1 protein (ab132404)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab34682 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
WB
Human
All applications
Rat
Chicken
Dog
Chimpanzee
Zebrafish
Application Abreviews Notes
WB (3)
1/500 - 1/2000. Detects a band of approximately 67 kDa (predicted molecular weight: 57 kDa). Splice variants exist for this protein that may result in the detection of lower molecular weight bands.
IHC-P
Use a concentration of 2 - 5 µg/ml.
ICC/IF
Use a concentration of 1 µg/ml.
Notes
WB
1/500 - 1/2000. Detects a band of approximately 67 kDa (predicted molecular weight: 57 kDa). Splice variants exist for this protein that may result in the detection of lower molecular weight bands.
IHC-P
Use a concentration of 2 - 5 µg/ml.
ICC/IF
Use a concentration of 1 µg/ml.

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.
  • Target information above from: UniProt accession Q16656 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 416677 Chicken
    • Entrez Gene: 4899 Human
    • Entrez Gene: 312195 Rat
    • Entrez Gene: 64604 Zebrafish
    • Omim: 600879 Human
    • SwissProt: Q16656 Human
    • SwissProt: Q62792 Rat
    • SwissProt: Q90X44 Zebrafish
    • Unigene: 654363 Human
    • Unigene: 224754 Rat
    • Unigene: 79816 Zebrafish
    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 (ab34682)
    Western blot - Anti-NRF1 antibody (ab34682)This image is courtesy of an anonymous Abreview
    All lanes : Anti-NRF1 antibody (ab34682) at 1/1000 dilution

    Lane 1 : Mouse liver whole tissue lysate
    Lane 2 : HeLa whole cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 57 kDa
    Observed band size: 55,67 kDa why is the actual band size different from the predicted?


    Exposure time: 50 seconds

    See Abreview

  • Western blot - Anti-NRF1 antibody (ab34682)
    Western blot - Anti-NRF1 antibody (ab34682)
    All lanes : Anti-NRF1 antibody (ab34682) at 1/500 dilution

    Lane 1 : HeLa Nuclear extract
    Lane 2 : HeLa Whole Cell lysate

    Secondary
    All lanes : IRDyeTM700 labelled goat-a-rabbit IgG at 1/2500 dilution

    Predicted band size: 57 kDa
    Observed band size: 67 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody (ab34682)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRF1 antibody (ab34682)
    Purified anti-NRF1 antibody (ab34682) was used at 5µg/ml to detect NRF1 in human testis. Nuclear staining in lymphocytes can be observed.
  • Immunocytochemistry/ Immunofluorescence - Anti-NRF1 antibody (ab34682)
    Immunocytochemistry/ Immunofluorescence - Anti-NRF1 antibody (ab34682)
    ICC/IF image of ab34682 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab34682, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (32)

Publishing research using ab34682? Please let us know so that we can cite the reference in this datasheet.

ab34682 has been referenced in 32 publications.

  • Hou Z  et al. Regulation of differential proton-coupled folate transporter gene expression in human tumors: transactivation by KLF15 with NRF-1 and the role of Sp1. Biochem J 476:1247-1266 (2019). PubMed: 30914440
  • Wang X  et al. Transcription regulation of NRF1 on StAR reduces testosterone synthesis in hypoxemic murine. J Steroid Biochem Mol Biol 191:105370 (2019). PubMed: 31028793
  • Kim D  et al. Protective effect of exercise training against the progression of Alzheimer's disease in 3xTg-AD mice. Behav Brain Res 374:112105 (2019). PubMed: 31325514
  • Senapati P  et al. Hyperinsulinemia promotes aberrant histone acetylation in triple-negative breast cancer. Epigenetics Chromatin 12:44 (2019). PubMed: 31315653
  • 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). PubMed: 31181625
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 10 Abreviews or Q&A

Western blot abreview for Anti-NRF1 antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Tissue lysate - whole (liver)
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted Dec 19 2014

Question

We have used ab34682 in the past and it has worked well; in this case our new vial of this antibody cross reacts with a pure protein that is totally unrelated to NRF1

Read More

Abcam community

Verified customer

Asked on Aug 23 2012

Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab34682.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Read More

Abcam Scientific Support

Answered on Aug 23 2012

Question

zur Abwechslung mal eine gute Nachricht. Der PGC1a AK (ab106814) scheint
sehr gut zu funktonieren auf dem Proteinlysat unserer Mausherzen. Anbei
ein Bild. Leider zeigt der Muskel nur eine sehr schawche Bande!?
Vielleicht können wir das nochmal mit mehr Proteinmenge versuchen!
Vielen Dank für ihre Hilfe und bis demnächst!
Beste Grüße,

Read More

Abcam community

Verified customer

Asked on Apr 19 2012

Answer

Vielen Dank für Ihre Nachricht! Es freut mich zu hören, dass das neue Röhrchen ein gutes Signal gibt auf dem Mausgewebe.

Es ist erstaunlich, dass das Signal auf dem Muskel nicht mehr da ist. In der Tat ist das Protein vielleicht auch degradiert? Haben Sie ein frisches Lysat, welches Sie testen könnten? Ist das Muskellysat auch von der Maus?

Ich hoffe, dass beim zweiten Versuch alles gut läuft. Ich würde mich über eine kurze Rückmeldung wieder freuen.

Vielen Dank für Ihre Kooperation! Ich freue mich wieder von Ihnen zu hören!

Read More

Abcam Scientific Support

Answered on Apr 19 2012

Question

follow up phone call

Read More

Abcam community

Verified customer

Asked on Apr 18 2012

Answer

Vielen Dank für Ihren Anruf.

Wie am Telefon besprochen, habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben mit einem neuen Röhrchen einer anderen Abpackung von ab34682.Die Bestellunghat die Referenznummer 1073393 und sollte morgen bei Ihnen ankommen.

Ich hoffe, dass dieses neue Röhrchen so funktioniert wie erwartet. Ich freue mich über eine kurze Rückmeldung Ihrerseits diesbezüglich. Bitte zögern Sie auch nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Read More

Abcam Scientific Support

Answered on Apr 18 2012

Question

wie letzte Woche telefonisch besprochen, haben wir die Hybridisierung mit
dem NRF1-AK (ab34682)wiederholt.
Leider konnten wir keine Bande auf der gewünschten Höhe detektieren. Das
Signal bei 70 kDa (siehe Anhang) kommt von der Hybridisierung mit einem AK
davor, der den gleichen 2.AK hat.
Vielen Dank für ihre Unterstützung. Mit besten Grüßen,

Read More

Abcam community

Verified customer

Asked on Apr 17 2012

Answer

Vielen Dank für Ihre Antwort.

Es tut mir leid, dass der ab34682 kein Signal mehr gibt. Leider kann ich mir nicht erklären, wieso dies so ist. Wir werden Ihnen gerne einen Ersatzantikörper senden. Könnten Sie bitte hierzu die Eckdaten des Protokolls mit dem angehängten Fragebogen eingeben (für unsereQualitätskontrolle), inklusive des Bestelldatums oder der Bestellnummer? Vielen Dank für Ihre Mitarbeit.

Bitte bestätigen Sie mir auch, ob Sie gerne ein Ersatzröhrchen von ab34682 hätten, oder lieber einen alternativen Antikörper.

Ich freue mich wieder von Ihnen zu hören.

Read More

Abcam Scientific Support

Answered on Apr 17 2012

Question

Thank you very much for your response. Your suggestion sounds good and it is acceptable to me.

Best regards,

Read More

Abcam community

Verified customer

Asked on Feb 27 2012

Answer

Thank you for your reply.

Please find the information below regarding the testing discount offer, to test ab90425 in ChIP. Also I have sent you ab90425 as a free of charge replacement for ab34682 which was mislabeled on our website. There should also be another email that you receive regarding the return instructions for the second vial of ab34682 to you purchased

Once again I am sorry about the confusion over the names on the website, but I hope that ab90425 will prove to be successful for you.


Testing Discount Details
DISCOUNT CODE: *********
Expiration date: **********

I am very pleased to hear you would like to accept our offer and test ab90524 in ChIP. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview forChIP 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: 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: www.abcam.com/collaborationdiscount.

Read More

Abcam Scientific Support

Answered on Feb 27 2012

Question

Thank you very much for your response. Your suggestion sounds good and it is acceptable to me.

Read More

Abcam community

Verified customer

Asked on Feb 27 2012

Answer

Thank you for contacting Abcam. Included in this email are the instructions for returning your item/s to Abcam. When you return the goods, please ensure that you follow the procedures as below:

- Return in the original Abcam box with the cooler box and ice pack (or similar) within 7 days from today.
- Return all documents (or copies of) that arrived with the product (if they are available – should be invoice, delivery note and datasheet)
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- Ship to the following address:

Attn: Returns ORDER REF: (***********)
Abcam Inc
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Bldg 200, Suite 341
Cambridge, MA 02139

There will be no restock fee on this order due to the error being on our end. The most up-to-date information regarding your order, invoice, and credit note is available online in your Abcam account information. If your order was placed by credit card, your credit card will be issued the refund amount. Please be advised that if your order was placed by Purchase Order, all customers have to submit a credit note with their invoice to their Accounts Payable department to be handled.

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Fax: 866 739 9884
Customer Service hours of operation: Monday-Friday 8:30 AM-9:00PM EST

Read More

Abcam Scientific Support

Answered on Feb 27 2012

Question

LOT NUMBER gr4790-3 ORDER NUMBER 901532 DESCRIPTION OF THE PROBLEM Multiple bands and the major band is wrong 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: rabbit •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: rabbit •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.

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Abcam community

Verified customer

Asked on Oct 04 2011

Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation. Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Abcam Scientific Support

Answered on Oct 04 2011

Western blot abreview for Anti-NRF1 antibody - ChIP Grade

Inconclusive
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (liver, muscle)
Loading amount
20 µg
Specification
liver, muscle
Gel Running Conditions
Reduced Denaturing (4-12% BioRad Criterion XT)
Blocking step
Milk 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 26 2009

Western blot abreview for Anti-NRF1 antibody - ChIP Grade

Below Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (C2C12)
Loading amount
15 µg
Specification
C2C12
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Jun 05 2009

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