Anti-Myc tag antibody [9E11] - ChIP Grade (ab56)

Reviews (7)Q&A (5)References (42)

Overview

  • Product name

    Anti-Myc tag antibody [9E11] - ChIP Grade
    See all Myc tag primary antibodies

  • Description

    Mouse monoclonal [9E11] to Myc tag - ChIP Grade

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt, ChIP, WB, IP, IHC-P, IHC-Frmore details

  • Species reactivity

    Reacts with: Chicken, Human, Saccharomyces cerevisiae

  • Immunogen

    Synthetic peptide corresponding to Human Myc tag aa 400-500 conjugated to keyhole limpet haemocyanin.

  • Positive control

  • General notes

    This antibody clone is manufactured by Abcam.

    ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab56 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
Flow Cyt 1/200.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 
ChIP Use at an assay dependent concentration. 2ul per 500ug of extract, see H. Zhang et al.
WB 1/500 - 1/1000. Predicted molecular weight: 49 kDa. Additional non-specific bands observed at 75, 110, 140 kDa using mouse and human cells (see Abreview).
IP Use a concentration of 5 µg/ml.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

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

  • Database links

    • Alternative names

      • c-myc tag antibody
      • Myc Epitope Tag antibody

    Images

    • Immunoprecipitation - Anti-Myc tag antibody [9E11] - ChIP Grade (ab56)
      Immunoprecipitation - Anti-Myc tag antibody [9E11] - ChIP Grade (ab56)

      Myc tag was immunoprecipitated using 0.5mg CHO overexpressing Stra8 whole cell lysate, 5µg of Mouse monoclonal to Myc tag and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

      The antibody was incubated under agitation with Protein G beads for 10min, CHO overexpressing Stra8 whole cell lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab56.

      Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

      Band: 49KDa; Myc tag

    • Western blot - Anti-Myc tag antibody [9E11] - ChIP Grade (ab56)
      Western blot - Anti-Myc tag antibody [9E11] - ChIP Grade (ab56)
      Anti-Myc tag antibody [9E11] - ChIP Grade (ab56) at 1/500 dilution + Recombinant Human c-Myc protein (ab84132) at 0.01 µg

      Secondary
      Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 49 kDa


      Exposure time: 4 minutes
    • Flow Cytometry - Anti-Myc tag antibody [9E11] - ChIP Grade (ab56)
      Flow Cytometry - Anti-Myc tag antibody [9E11] - ChIP Grade (ab56)

      Overlay histogram showing HL60 cells stained with ab56 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    • Western blot - Anti-Myc tag antibody [9E11] - ChIP Grade (ab56)
      Western blot - Anti-Myc tag antibody [9E11] - ChIP Grade (ab56)
      All lanes : Anti-Myc tag antibody [9E11] - ChIP Grade (ab56) at 1 µg/ml

      Lane 1 : HEK293 cell lysate at 20 µg
      Lane 2 : Recombinant Human c-Myc protein (ab84132) at 0.1 µg
      Lane 3 : E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395) at 0.1 µg

      Secondary
      All lanes : Goat polyclonal to Mouse IgG H&L Pre-Adsorbed (HRP) at 1/5000 dilution

      Predicted band size: 49 kDa

    Protocols

    References

    This product has been referenced in:

    • Shang Z  et al. LncRNA PCAT1 activates AKT and NF-?B signaling in castration-resistant prostate cancer by regulating the PHLPP/FKBP51/IKKa complex. Nucleic Acids Res N/A:N/A (2019). Read more (PubMed: 30773595) »
    • Wong M  et al. JMJD6 is a tumorigenic factor and therapeutic target in neuroblastoma. Nat Commun 10:3319 (2019). Read more (PubMed: 31346162) »
    See all 42 Publications for this product

    Publishing research using ab56? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 12 Abreviews or Q&A

    Western blot abreview for Anti-c-Myc antibody [9E11] - ChIP Grade

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (4-12% Nupage gel)
    Sample
    Human Cell lysate - whole cell (HEK 293)
    Specification
    HEK 293
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 26 2014

    Western blot abreview for Anti-c-Myc antibody [9E11] - ChIP Grade

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MCF7)
    Loading amount
    10 µg
    Specification
    MCF7
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 26 2011

    ChIP abreview for Anti-c-Myc antibody [9E11] - ChIP Grade

     Good
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (skin fibroblasts)
    Specification
    skin fibroblasts
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: Formaldehyde
    Detection step
    Real-time PCR
    Positive control
    NPM1 cMYC binding site
    Negative control
    IgG pull-down
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 15 2011

    Western blot abreview for Anti-c-Myc antibody [9E11] - ChIP Grade

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (huh-7)
    Loading amount
    15 µg
    Specification
    huh-7
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 08 2010

    Western blot abreview for Anti-c-Myc antibody [9E11] - ChIP Grade

     Average
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (mammary epithelial)
    Loading amount
    80 µg
    Specification
    mammary epithelial
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 25 2009

    Western blot abreview for Anti-c-Myc antibody [9E11] - ChIP Grade

     Below Average
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Breast cancer cell-lines)
    Loading amount
    20 µg
    Specification
    Breast cancer cell-lines
    Gel Running Conditions
    Non-reduced Denaturing (4-20% Nupage gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Dr. Cherie Blenkiron

    Verified customer

    Submitted Jul 18 2008

    Question

    I'd like to know whether this antibody works efficiently in ChIP (conventional paraformaldehyde fixation) of Myc-tagged (one tag) proteins expressed in mammalian tissue culture cells. .

    Read More
    Answer

    This antibody was qualified as "ChIP grade" following its application in the following publications; Robert F et al. Global position and recruitment of HATs and HDACs in the yeast genome. Mol Cell 16:199-209 (2004). PubMed: 15494307 Zhang H et al. The Yaf9 component of the SWR1 and NuA4 complexes is required for proper gene expression, histone H4 acetylation, and Htz1 replacement near telomeres. Mol Cell Biol 24:9424-36 (2004). PubMed: 15485911 We have received reports suggesting that the antibody does not give a huge enrichment but that the enrichment is specific. Furthermore it is guaranteed for this purpose. I would recommend that you consult the publications for details of the approach that was adopted. From my recollection they did use proteins tagged with more than one Myc-tag.

    Read More

    Western blot abreview for Anti-c-Myc antibody [9E11] - ChIP Grade

     Good
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Pancreatic cell lines)
    Specification
    Pancreatic cell lines
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Dr. Lynda Elghazi-Cras

    Verified customer

    Submitted Jul 25 2005

    Question

    I would like to know which of your human breast tumor slides would be the best positive control for antibody ab56.

    Read More
    Answer

    Thank you for your enquiry and your interest in our products. I would suggest taking a look at the published references to have some more information about the breast cancer cell types in which c-myc is expressed. Locker AP et al. c-myc oncogene product expression and prognosis in operable breast cancer. Br J Cancer 60:669-72 (1989). PubMed: 2679850 Once you know which tumour types you are looking for, you can conduct a search in our on-line catalogue. We have several human breast tissue slides such as: ab4696: fibroadenoma, ab4701: adenocarcinoma, ab4697: dusctal carcinoma, ab4698: lobular carcinoma, ab4700: medullary carcinoma

    Read More

    Question

    Can I use c-myc 9e11 for histochemical localization in paraffin embedded sections and what antigen retrival steps do I take and what conc is optimum??

    Read More
    Answer

    Thsi antibody works well for histochemical localization in paraffin embedded sections. We would recommend a microwave retrieval (10 minutes in 0.01M citrate buffer, pH 6.0, the specific time will depend on the power of the microwave). Leave the 1ary antibody on overnight at a concentration of 5ug/ml.

    Read More

    1-10 of 12 Abreviews or Q&A

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