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

41-50 of 51 Abreviews or Q&A

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.

Read More
Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (Cos7 transfected with pCMV-Nle-Myc)
Loading amount
1e+006 cells
Specification
Cos7 transfected with pCMV-Nle-Myc
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted May 05 2006

Application
Immunocytochemistry/ Immunofluorescence
Sample
African Green Monkey Cell (Cos7 transfected with pCMV-Nle-Myc)
Specification
Cos7 transfected with pCMV-Nle-Myc
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted May 05 2006

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293 FT)
Total protein in input
500 µg
Specification
293 FT
Immuno-precipitation step
Protein A/G

Abcam user community

Verified customer

Submitted Apr 25 2006

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (embryonic kidney)
Specification
embryonic kidney
Immuno-precipitation step
Protein A

Ms. Michelle Badura

Verified customer

Submitted Mar 14 2006

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.

Read More
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293-T cells)
Specification
293-T cells
Immuno-precipitation step
Protein G

Dr. Mary Donaldson

Verified customer

Submitted Oct 19 2005

Application
Immunoprecipitation
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (asynchronous population)
Specification
asynchronous population
Immuno-precipitation step
Protein A

Mr. Mark Chee

Verified customer

Submitted Aug 31 2005

Answer

Thank you for your enquiry and patience. The originator of ab9106 has informed me that the antibody was tested in IHC-frozen sections at a dilution of 1:500. We don't have a specific protocol available, but would recommend referring to our IHC-frozen sections protocol which can be found by clicking on the protocols tab located on the online datasheet for ab9106. If you have any additional questions, please contact us again.

Read More

41-50 of 51 Abreviews or Q&A

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