製品の概要

  • 製品名

    Anti-Myc tag antibody [9E11] - ChIP Grade
    Myc tag 一次抗体 製品一覧
  • 製品の詳細

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

    Mouse
  • アプリケーション

    適用あり: Flow Cyt, ChIP, WB, IP, IHC-P, IHC-Frmore details
  • 種交差性

    交差種: Chicken, Human, Saccharomyces cerevisiae
  • 免疫原

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

  • ポジティブ・コントロール

  • 特記事項

    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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 関連性

    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.
  • 細胞内局在

    Nuclear
  • 参照データベース

  • 別名

    • c-myc tag antibody
    • Myc Epitope Tag antibody

画像

  • 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

  • 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
  • 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.

  • 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

プロトコール

参考文献

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) »
  • Ma Q  et al. Regulation of the MAPK signaling pathway by miR-421-5p in rats under light pollution. Int J Mol Med 42:3329-3343 (2018). Read more (PubMed: 30221682) »
See all 38 Publications for this product

レビューと Q&A

1-10 of 12 Abreviews or Q&A

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

Abcam user community

Verified customer

投稿 Sep 26 2014

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

Abcam user community

Verified customer

投稿 Oct 26 2011

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

Abcam user community

Verified customer

投稿 Jul 15 2011

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

Abcam user community

Verified customer

投稿 Jun 08 2010

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

Abcam user community

Verified customer

投稿 Jun 25 2009

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

Dr. Cherie Blenkiron

Verified customer

投稿 Jul 18 2008

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
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%

Dr. Lynda Elghazi-Cras

Verified customer

投稿 Jul 25 2005

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

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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