Anti-Myc tag antibody (ab9106)

Rabbit polyclonal Myc tag antibody. Validated in WB, IP, IHC, ICC, Flow Cyt, EM, ICC/IF. Cited in 255 publication(s). Independently reviewed in 38 review(s).

Overview

  • Product name
  • Description
    Rabbit polyclonal to Myc tag
  • Host species
    Rabbit
  • Specificity
    ELISA: Antibody specificity was verified by ELISA against the peptide (EQKLISEEDL). A 1:60,000 dilution of the antibody gave an O.D.=1.0 in a 15 minute reaction using HRP-conjugated Goat Anti Rabbit IgG at 1:20,000 and TMB as the substrate. Appropriate specificity controls were run.
  • Tested applications
    Suitable for: IHC-Fr, IP, WB, IHC-P, Flow Cyt, ICC/IF, ICC, Electron Microscopymore details
  • Immunogen

    Synthetic peptide corresponding to Myc tag conjugated to keyhole limpet haemocyanin.
    Sequence:

    EQKLISEEDL


    (Peptide available as ab13837)

  • Positive control
    • ICC/IF: HeLa cells transfected with a Myc-tagged membrane targeting protein. IP: Huh7 cells expressing myc-tagged CDC42 protein. Human 293FT cells co-transfected with myc tagged cdc25 along with an HA tagged viral protein which binds to cdc25.
  • General notes

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

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: 0.1% Sodium azide
  • Concentration information loading...
  • Purity
    Affinity purified
  • Purification notes
    Antibodies were immunoaffinity purified using the peptide immobilized on a solid phase. Antibody concentration was determined by extinction coefficient : O.D. 1.4 at 280nM equals 1.0 mg of IgG.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab9106 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
IHC-Fr 1/500.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Can be blocked with Human c-Myc peptide (ab13837).
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration. PubMed: 19793804

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration. PubMed: 16093346
ICC Use at an assay dependent concentration.
Electron Microscopy Use at an assay dependent concentration. See Abreview for further details (submitted by Eeva-Liisa Eskelinen).

Target

  • Relevance
    Epitope tags are short peptide sequences that are easily recognized by tag-specific antibodies. Due to their small size, epitope tags do not affect the tagged protein’s biochemical properties. Most often sequences encoding the epitope tag are included with target DNA at the time of cloning to produce fusion proteins containing the epitope tag sequence. This allows anti-epitope tag antibodies to serve as universal detection reagents for any tag containing protein produced by recombinant means. This means that anti-epitope tag antibodies are a useful alternative to generating specific antibodies to identify, immunoprecipitate or immunoaffinity purify a recombinant protein. The anti-epitope tag antibody is usually functional in a variety of antibody-dependent experimental procedures. Expression vectors producing epitope tag fusion proteins are available for a variety of host expression systems including bacteria, yeast, insect and mammalian cells.
  • Cellular localization
    Nuclear
  • Alternative names
    • c-myc tag antibody
    • Myc Epitope Tag antibody

Images

  • Y. enterocolitica ΔHOPEMT carrying plasmids encoding Myc-tagged CT082 or CT695, and HA-tagged Slc1 (as indicated) were grown in non-secreting conditions. Chloramphenicol was added (time?=?0 min) to stop bacterial protein synthesis and samples were taken at the indicated time points. Samples were analyzed by immunoblotting using ab9106, anti-HA, and anti-TEM antibodies.

  • ab9106 staining the Myc tag in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells transfected with a Myc-tagged membrane targeting protein by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with paraformaldehyde, permeabilized with Triton X (0.5%) and blocked with 3% BSA for 1 hour at 24°C. Samples were incubated with primary antibody (1/500) for 1 hour at 24°C. An Alexa Fluor®488-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

  • ab9106 at 4 µg/mg lysate used in Huh7 (Human cell line) whole cell lysate (1x106 cells).

    Cells expressed myc-tagged CDC42 protein. Immunoprecipitation step performed using Protein A matrix. Incubation time 4 hours at 4°C. Western Blot antibody used at 1/300 dilution.

    See Abreview

  • ab9106 used at 1/250 in the immunoprecipitation of transfected human 293FT cells (whole cell lysate).

    In this experiment cells were co-transfected with myc tagged cdc25 along with an HA tagged viral protein which binds to cdc25. The cell lysate was immunoprecipitated using ab9106, run on a 10% gel. The blot was probed for HA and HA tagged protein (which co-IPed with Myc tagged protein) and this is seen at ~15KDa.

    Lane 1: Input non-transfected cells

    Lane 2: Immunoprecipitated non-transfected cells

    Lane 3: Input co-transfected cells

    Lane 4: Immunoprecipitated co-transfected cells

    This image is courtesy of an Abreview submitted on 23 March 2006.

    See Abreview

  • ab9106 at 1/2000 staining mouse embryonic fibroblasts expressing myc-tagged protein by ICC.

    The cells were paraformaldehyde fixed and blocked with 3% BSA prior to incubation with the antibody for 1 hour. A goat anti-rabbit IgG antibody (10 nm gold conjugated) was used as the secondary.

    The image shown is immuno electron microscopical staining with thawed cryosections.

    See Abreview

References

This product has been referenced in:
  • Jin Y  et al. TRIM21 mediates ubiquitination of Snail and modulates epithelial to mesenchymal transition in breast cancer cells. Int J Biol Macromol 124:846-853 (2019). Read more (PubMed: 30502437) »
  • Safiulina D  et al. Miro proteins prime mitochondria for Parkin translocation and mitophagy. EMBO J 38:N/A (2019). Read more (PubMed: 30504269) »
See all 258 Publications for this product

Customer reviews and Q&As

1-10 of 13 Q&A

Answer

Thank you for your email.

Unfortunately the delay wasn't from Abcam, we have shipped the product at correct temperature. Please check your delivery service and let them know these products are temperature sensitive and they need to be delivered without delays.

We will be shipping a new vial as replacement to the next order of Nulife. Please make sure the customer gets this vial without delays. The new order number is 1229622.

I hope this information will be helpful. Should you have any question please do not hesitate to contact me.

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Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

- Antibodies usually are OK for about a week at room temperature which however varies from product to product so for how long this antibody was delayed in shipment?
- How do you define the problem? Is it no band, high background or multiple band etc?
- The antibody is against myc tag so have you used the antibody against protein of interest? How were the results?
- Could you provide an image?

Thank you very much for your cooperation. I will look forward to hearing from you soon.

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Answer

Thank you for contacting us.

From the looks of it, this is not a suitable antibody for you, as it is only suitable for the tag (only a few AA long)but not for the c-Myc protein (48 kDa). I checked the sequence and it looks like this antibody coulddetect c-Myc, but there may be a lot of other cross-reactivity as the sequence is so short.Fortunately, we have a few c-Myc antibodies in our catalogue. I picked three antibodies with the best ratings for your applications and the most publications:

https://www.abcam.com/index.html?datasheet=32 (or use the following: https://www.abcam.com/index.html?datasheet=32).



https://www.abcam.com/index.html?datasheet=39688 (or use the following: https://www.abcam.com/index.html?datasheet=39688).

https://www.abcam.com/index.html?datasheet=32072 (or use the following: https://www.abcam.com/index.html?datasheet=32072).



Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

DISCOUNT CODE: ***
Expiration date: June 17th, 2012
I am very pleased to hear you would like to accept our offer and test ab9106 in ChIP. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ChIP 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 getting back to me and for providing some further information about your research project.

Currently, we have 2 monoclonal antibodies against Myc tag: ab18185 and ab94936. Although they are from the same clone but from two sources and the testing conditions were different. ab94936 reacts with recombinant fragment so the other antibody ab18185 may suit your need better.

There are two specific publication for ab18185 you could take a look at to get some more information:

Biswas Ket al. A comprehensive functional characterization of BRCA2 variants associated with Fanconi anemia using mouse ES cell-based assay. Blood 118:2430-42 (2011). WB.http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=21719596&dopt=Abstract
Ohe Met al. Emi2 inhibition of the anaphase-promoting complex/cyclosome absolutely requires Emi2 binding via the C-terminal RL tail. Mol Biol Cell 21:905-13 (2010). WB.http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=20089832&dopt=Abstract



If you need any further assistance in the future, please do not hesitate to contact me.

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Answer

Thank you for your enquiry and interest in our products.

Could you please let me know if you wish to detect a Myc tagged protein or myc itself? For what application you are looking for the alternative antibodies and what type of samples you are using? Is there any reason why you wish to switch from ab9106 to another antibody? Any additional information you could provide would help me advise some alternative products for you.

I look forward to hearing from you soon.

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Answer

Thank you for contacting Abcam regarding our Anti-myc antibodies. I have confirmed that yes, ab1326 is the conjugated version of ab9106. I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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Answer

I'm afraid I have received very little information from the source of ab9106. I was informed just a few minutes ago " a customer of mine had sent me the anti-Myc image, but only provided me with the working dilution. I have been unable to track them down at their university". I am therefore unable to find out what type of fixative was used. For ab32, the information that it works in IF dates from 2001, where a customer told us it worked well for him. At the time we did not request further details and therefore unfortunately again we do not know if 4%PFA was the fixative successfully used. If you are able to try different fixatives on your cells I would therefore recommend to try both 4% PFA and acetone as fixatives and see which one works well for you. If you cannot have your cells fixed in anything other than 4% PFA I would recommend not to purchase these products as I'm not sure they will work with 4% PFA. I would be happy to enquire if ab9132 will work on 4% PFA fixed cells, however the detection method was chromogenic, not fluorescent, therefore the antibody may or may not work in IF, we don't know at this stage. I'm sorry I cannot help you more on this occasion,

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Question
Answer

Thank you for your enquiry. The comparison of the amino acid of the sequence used to generate this antibody with the full-length precursor of mouse myc (primary accession number P01108) shows 70% identity. The file for this comparison can be found at this link for the next 24 hours: http://www.ebi.ac.uk/cgi-bin/clustalw/result?tool=clustalw&jobid=clustalw-20060922-16195297&poll=yes Recognition of mouse myc by this antibody, given the 70% identity, is difficult to predict. However, two of the amino acid pairs consist of biochemically similar amino acids(designated by the ":" symbol in this alignment tool), and the antibody is polyclonal, so it will potentially recognize more than one epitope on the sequence. If you decide to try the antibody to detect endogenous mouse myc, we would appreciate your feedback. We have a rewards incentive for reviews of our antibodies called Abpoints, the details of which can be found on our website. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your enquiry. This antibody was tested for quality control by loading 50 ng of tagged protein per well. I think it would detect lower concentrations but I would suggest a starting dilution of 2 ug/ml of antibody and increase the normal incubation period from 60 min to 120 min. I hope this information helps. Please do not hesitate to contact us if you need anything further.

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1-10 of 13 Q&A

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