Overview

  • Product name
  • 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
    Suitable for: ChIP, WB, IHC-P, ICC/IF, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Dog, Chimpanzee, Zebrafish
  • Immunogen

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

  • Positive control
    • HeLa Whole Cell Lysate HeLa Cell Nuclear Extract Human testis

Properties

Applications

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.

Application Abreviews Notes
ChIP Use at an assay dependent concentration.
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.
ELISA 1/3000 - 1/10000.
EMSA Use at an assay dependent concentration. PubMed: 21609478

Target

Images

  • 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

  • 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?

  • Purified anti-NRF1 antibody (ab34682) was used at 5µg/ml to detect NRF1 in human testis. Nuclear staining in lymphocytes can be observed.
  • 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.

References

This product has been referenced in:
  • Yu B  et al. PGC-1a Controls Skeletal Stem Cell Fate and Bone-Fat Balance in Osteoporosis and Skeletal Aging by Inducing TAZ. Cell Stem Cell 23:193-209.e5 (2018). Read more (PubMed: 30017591) »
  • Zhang T  et al. A negative feedback loop between microRNA-378 and Nrf1 promotes the development of hepatosteatosis in mice treated with a high fat diet. Metabolism 85:183-191 (2018). ChIP . Read more (PubMed: 29625129) »
See all 23 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Dec 19 2014

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.

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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!

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Question
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.

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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.

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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.

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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)
- Free shipping: Please return via FEDEX PRIORITY OVERNIGHT SHIPPING and use the Fedex account number - **********.
- Ship to the following address:

Attn: Returns ORDER REF: (***********)
Abcam Inc
One Kendall Square
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.

We apologize for this inconvenience you may have been caused by this error. If you have any other questions please do not hesitate to contact us.

Email: US.orders@Abcam.com
Phone: 888-77-ABCAM(22226)
Fax: 866 739 9884
Customer Service hours of operation: Monday-Friday 8:30 AM-9:00PM EST

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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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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.

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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

Abcam user community

Verified customer

Submitted Oct 26 2009

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

Abcam user community

Verified customer

Submitted Jun 05 2009

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

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