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

Overview

  • Product nameAnti-c-Myc antibody [9E11] - ChIP Grade
    See all c-Myc primary antibodies
  • Description
    Mouse monoclonal [9E11] to c-Myc - ChIP Grade
  • Conjugation notes-
  • Tested applicationsFlow Cyt, ChIP, WB, IP, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Chicken, Human, Saccharomyces cerevisiae
  • Immunogen

    Synthetic peptide:

    AEEQKLISEEDL

    conjugated to KLH, corresponding to amino acids 408-420 of Human c-Myc.

  • Positive control
  • General notes

    This product is available conjugated to FITC see ab150238

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.
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

  • FunctionParticipates in the regulation of gene transcription. Binds DNA in a non-specific manner, yet also specifically recognizes the core sequence 5'-CAC[GA]TG-3'. Seems to activate the transcription of growth-related genes.
  • Involvement in diseaseNote=Overexpression of MYC is implicated in the etiology of a variety of hematopoietic tumors.
    Note=A chromosomal aberration involving MYC may be a cause of a form of B-cell chronic lymphocytic leukemia. Translocation t(8;12)(q24;q22) with BTG1.
    Defects in MYC are a cause of Burkitt lymphoma (BL) [MIM:113970]. A form of undifferentiated malignant lymphoma commonly manifested as a large osteolytic lesion in the jaw or as an abdominal mass. Note=Chromosomal aberrations involving MYC are usually found in Burkitt lymphoma. Translocations t(8;14), t(8;22) or t(2;8) which juxtapose MYC to one of the heavy or light chain immunoglobulin gene loci.
  • Sequence similaritiesContains 1 basic helix-loop-helix (bHLH) domain.
  • Post-translational
    modifications
    Phosphorylated by PRKDC. Phosphorylation at Thr-58 and Ser-62 by GSK3 is required for ubiquitination and degradation by the proteasome.
    Ubiquitinated by the SCF(FBXW7) complex when phosphorylated at Thr-58 and Ser-62, leading to its degradation by the proteasome. In the nucleoplasm, ubiquitination is counteracted by USP28, which interacts with isoform 1 of FBXW7 (FBW7alpha), leading to its deubiquitination and preventing degradation. In the nucleolus, however, ubiquitination is not counteracted by USP28, due to the lack of interaction between isoform 4 of FBXW7 (FBW7gamma) and USP28, explaining the selective MYC degradation in the nucleolus. Also polyubiquitinated by the DCX(TRUSS) complex.
  • Cellular localizationNucleus > nucleoplasm. Nucleus > nucleolus.
  • Information by UniProt
  • Database links
  • Formc-Myc is also expressed in the cytoplasm.
  • Alternative names
    • AU016757 antibody
    • Avian myelocytomatosis viral oncogene homolog antibody
    • bHLHe39 antibody
    • c Myc antibody
    • Cellular myelocytomatosis oncogene antibody
    • Class E basic helix-loop-helix protein 39 antibody
    • MGC105490 antibody
    • MRTL antibody
    • Myc antibody
    • Myc protein antibody
    • Myc proto oncogene protein antibody
    • Myc proto-oncogene protein antibody
    • Myc-related translation/localization regulatory factor antibody
    • MYC_HUMAN antibody
    • Myc2 antibody
    • myca antibody
    • MYCC antibody
    • Myelocytomatosis oncogene a antibody
    • Myelocytomatosis oncogene antibody
    • Niard antibody
    • Nird antibody
    • Oncogene Myc antibody
    • OTTHUMP00000158589 antibody
    • Proto-oncogene c-Myc antibody
    • Protooncogene homologous to myelocytomatosis virus antibody
    • RNCMYC antibody
    • Transcription factor p64 antibody
    • Transcriptional regulator Myc-A antibody
    • v myc avian myelocytomatosis viral oncogene homolog antibody
    • v myc myelocytomatosis viral oncogene homolog (avian) antibody
    • V-Myc avian myelocytomatosis viral oncogene homolog antibody
    • v-myc myelocytomatosis viral oncogene homolog (avian) antibody
    • zc-myc antibody
    see all

Anti-c-Myc antibody [9E11] - ChIP Grade images

  • c-Myc was immunoprecipitated using 0.5mg CHO overexpressing Stra8 whole cell lysate, 5µg of Mouse monoclonal to c-Myc 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; c-Myc

  • 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.

  • Anti-c-Myc antibody [9E11] - ChIP Grade (ab56) at 1/1000 dilution + whole cell lysate prepared from huh-7 cells at 15 µg

    Secondary
    Anti-mouse IgG conjugated to HRP at 1/10000 dilution

    Predicted band size : 49 kDa

    Image courtesy of an anonymous Abreview.


  • developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 49 kDa

References for Anti-c-Myc antibody [9E11] - ChIP Grade (ab56)

This product has been referenced in:
  • Sun P  et al. Pygo2 regulates ß-catenin-induced activation of hair follicle stem/progenitor cells and skin hyperplasia. Proc Natl Acad Sci U S A 111:10215-20 (2014). Read more (PubMed: 24982158) »
  • Wright PW  et al. Characterization of a weakly expressed KIR2DL1 variant reveals a novel upstream promoter that controls KIR expression. Genes Immun N/A:N/A (2014). Read more (PubMed: 24989671) »

See all 19 Publications for this product

Product Wall

Thank you for taking time to contact us with these details. I am sorry to hear that this antibody is not providing satisfactory results.
We firstly would appreciate to receive more details on the experimental procedure. I have therefore attached a...

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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
Username

Abcam user community

Verified customer

Submitted Sep 26 2014

According to our records, ab56 was proving difficult to use in ChIP and we were in contact in order to help resolve the issue.
Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the di...

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Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Oct 26 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Jul 15 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Jun 08 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted 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
Username

Dr. Cherie Blenkiron

Verified customer

Submitted Jul 18 2008

Thank you for your enquiry. 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). Pu...

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Thank you for your enquiry. At Abcam we have validated a panel of epitope tagged antibodies for use in chromatin immunoprecipitation (see link at the bottom of this email). I would like to recommend the following antibodies in view of the tags that...

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