Anti-c-Myc [9E10] antibody - ChIP Grade (ab32)

Overview

  • Product nameAnti-c-Myc [9E10] antibody - ChIP GradeSee all c-Myc primary antibodies ...
  • Description
    Mouse monoclonal [9E10] to c-Myc - ChIP Grade
  • Tested applicationsICC/IF, ICC, ChIP, IHC (Methanol fixed), Flow Cyt, WB, IP, ELISA, IHC-P, IHC-Fr, Purification more details
  • Species reactivity
    Reacts with: Mouse, Human, Fruit fly (Drosophila melanogaster)
  • Immunogen

    Synthetic peptide: AEEQKLISEE DLLRKRREQL KHKLE conjugated to KLH, corresponding to C terminal amino acids 408-432 of Human c-Myc.

  • Epitopeaa 410-419 of human c-Myc.
  • Positive control

Properties

Applications

Our Abpromise guarantee covers the use of ab32 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
ICC/IF Use a concentration of 5 µg/ml.
ICC 1/2000.

See Abreviews.

ChIP Use at an assay dependent concentration.
IHC (Methanol fixed) 1/200. PubMed: 17329357
Flow Cyt Use 1µg for 106 cells.
WB 1/500 - 1/1000. This antibody has been used for immunolocalisation with gold conjugated secondaries on plant tissue.
IP Use at 6 µg/mg of lysate. It is not known whether this antibody is suitable for immunoprecipitation of native c-myc protein.
ELISA Use at an assay dependent concentration.
IHC-P 1/250 - 1/500.
IHC-Fr 1/1000. See Abreviews.
Purification 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.
  • Target information above from: UniProt accession P01106 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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
    • c Myc Epitope Tag antibody
    • c Myc Tag antibody
    • Class E basic helix-loop-helix protein 39 antibody
    • MRTL antibody
    • Myc antibody
    • Myc Epitope Tag 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
    • MYCC 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
    see all

Anti-c-Myc [9E10] antibody - ChIP Grade images

  • Anti-c-Myc [9E10] antibody - ChIP Grade (ab32) at 1 µg/ml + E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395) at 10 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 41 kDa
    Observed band size : 45 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • Overlay histogram showing Jurkat cells stained with ab32 (red line). The cells were fixed with 80% methanol (5 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 (ab32, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.



  • Predicted band size : 41 kDa


    Phosphorylation of Cdc15 changes during the cell cycle. Exponentially growing cells (cyc) of CDC15-MYC9 (W1114) were arrested in G1 with a factor pheromone (a) and released into fresh medium at 258C. Cells were harvested at the indicated times, the percentage of divided nuclei was determined by DAPI staining of fixed cells,and proteins were analyzed by Western blotting with 9E10 (ab32).

    Menssen R et al., (2001) Curr Biol. Mar 6;11(5):345-50.

  • ICC/IF image of ab32 stained human Hek293 cells. The cells were methanol fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab32, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • ab32 staining c-Myc in HEK293 cells transfected with pCMV4-myc-Mfn2 by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with paraformaldehyde, permeabilized with 1% Triton X-100 and blocked with 5% horse serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/1200 in 5% horse serum) for 1 hour. An AlexaFluor®488-conjugated goat anti-mouse polyclonal IgG (1/400) was used as the secondary antibody.

    See Abreview

Protocols

References for Anti-c-Myc [9E10] antibody - ChIP Grade (ab32)

This product has been referenced in:
  • Alimova I  et al. Inhibition of EZH2 suppresses self-renewal and induces radiation sensitivity in atypical rhabdoid teratoid tumor cells. Neuro Oncol 15:149-60 (2013). Read more (PubMed: 23190500) »
  • Raynaud F  et al. Shank3-Rich2 interaction regulates AMPA receptor recycling and synaptic long-term potentiation. J Neurosci 33:9699-715 (2013). Mouse . Read more (PubMed: 23739967) »

See all 47 Publications for this product

Product Wall

Application Immunoprecipitation
Immuno-precipitation step Other - Magnetic Protein G
Sample Human Cell lysate - whole cell (HeLa)
Specification HeLa
Total protein in input 20 µg
Username

Abcam user community

Verified customer

Submitted Jul 15 2013

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (6%)
Sample Human Cell lysate - whole cell (HeLa)
Specification HeLa
Blocking step BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jul 04 2013


The antibody was originally designed for pull down (IP) and ChIP of c-myc-tagged proteins. And that is how it has been used by other customers who submitted IP and ChIP reviews.

The information that "This antibody is NOT suitable for...

Read More

Thank you for contacting us.


The alternatives are as follows

1. ab103746; http://www.abcam.com/BrightMount-25ml-Aqueous-Mounting-Medium-for-Fluorescent-Staining-ab103746.html

or ab128982; http://www.abcam.com/Aqueous-Mou...

Read More
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HEK293)
Specification HEK293
Fixative Paraformaldehyde
Permeabilization Yes - 1%triton x100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Sep 04 2012

Application Western blot
Sample Human Cell lysate - whole cell (HEK293 CELL)
Loading amount 20 µg
Specification HEK293 CELL
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Aug 22 2012

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (HEK293)
Total protein in input 500 µg
Specification HEK293
Immuno-precipitation step Other - invitrogen IgG Dynabeads
Username

Abcam user community

Verified customer

Submitted Aug 21 2012

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has c...

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Application Western blot
Sample Human Cell lysate - whole cell (H1792 lung cancer cell line)
Loading amount 10 µg
Specification H1792 lung cancer cell line
Gel Running Conditions Reduced Denaturing (4-12% bis-Tris Nupage gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 16 2012

We unfortunately do not have new lot for this antibody yet. However I can send you a vial which was recently aliquoted from the same mother lot. Would the customer like to try this new vial?
Thank you very much for your cooperation in this case.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"