Anti-c-Myc [9E10] antibody - ChIP Grade (ab32)
- Product nameAnti-c-Myc [9E10] antibody - ChIP GradeSee all c-Myc primary antibodies ...
- DescriptionMouse 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 reactivityReacts with: Mouse, Human, Fruit fly (Drosophila melanogaster)
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
- Purchase matching WB positive control:Active human c-Myc full length protein
- HL-60 cells. Breast tumors. Flow Cyt: This antibody gave a positive signal in methanol fixed/Tween permeabilsed Jurkat cells.
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferpH: 7.4
Preservative: 0.02% Sodium azide
Contains 0.4M Arginine
- Concentration information loading...
- PurityIgG fraction
- Clonality Monoclonal
- Clone number9E10
- Light chain typekappa
- Epigenetics and Nuclear Signaling
- Domain Families
- HLH / Leucine Zipper
- HLH / Leucine Zipper
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.
|ICC/IF||ICC/IF: 1/2000. See Abreviews.|
|ChIP||ChIP: Use at an assay dependent dilution.|
|IHC (Methanol fixed)||IHC (Methanol fixed): 1/200. PubMed: 17329357|
|Flow Cyt||Flow Cyt: Use 1µg for 106 cells.|
|WB||WB: 1/500 - 1/1000. This antibody has been used for immunolocalisation with gold conjugated secondaries on plant tissue.|
|IP||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||ELISA: Use at an assay dependent dilution.|
|IHC-P||IHC-P: 1/250 - 1/500.|
|IHC-Fr||IHC-Fr: 1/1000. See Abreviews.|
|Purification||P: Use at an assay dependent dilution.|
- 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.
modificationsPhosphorylated 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.
- Formc-Myc is also expressed in the cytoplasm.
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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
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.
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) »