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

Overview

• Product nameAnti-c-Myc antibody [9E10] - ChIP GradeSee all c-Myc primary antibodies ...
• Description
  Mouse monoclonal [9E10] to c-Myc - ChIP Grade
• SpecificityThis anti-c-Myc antibody recognises recombinant proteins incorporating a c-Myc epitope tag. It also detects human c-Myc protein and a peptide in random coil configuration not as a helix. It does not react with v-myc.
• 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 controlHL-60 cells. Breast tumors. Flow Cyt: This antibody gave a positive signal in methanol fixed/Tween permeabilsed Jurkat cells.

Properties

• FormLiquid
• 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 bufferPreservative: 0.02% Sodium Azide
  Constituents: PBS
• Concentration information loading...
• PurityIgG fraction
• Clonality Monoclonal
• Clone number9E10
• MyelomaSp2/0
• IsotypeIgG1
• Light chain typekappa
• Research Areas
  • Stem Cells
  • Signaling Pathways
  • TGF beta
  • Nuclear

  • Cancer
  • Tumor biomarkers
  • Oncoproteins

  • Cancer
  • Cell cycle
  • Cell differentiation

  • Cancer
  • Oncoproteins/suppressors
  • Oncoproteins
  • Transcription factors

  • Tags & Cell Markers
  • Epitope Tags
  • Myc Tag

  • Epigenetics and Nuclear Signaling
  • ChIP'ing antibodies
  • ChIP'ing antibodies

  • Epigenetics and Nuclear Signaling
  • Transcription
  • Transcription Factors

  • Epigenetics and Nuclear Signaling
  • Transcription
  • Domain Families
  • HLH / Leucine Zipper
  • HLH / Leucine Zipper

Applications

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
  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.

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
  • Entrez Gene: 4609 Human
  • Entrez Gene: 17869 Mouse
  • Omim: 190080 Human
  • SwissProt: P01106 Human
  • SwissProt: P01108 Mouse
  • Unigene: 202453 Human
  • Unigene: 2444 Mouse

• Alternative names
  AU016757 antibodyAvian myelocytomatosis viral oncogene homolog antibodybHLHe39 antibody

  c Myc antibodyc Myc Epitope Tag antibodyc Myc Tag antibodyCellular myelocytomatosis oncogene antibodyClass E basic helix-loop-helix protein 39 antibodyMGC105490 antibodyMRTL antibodyMyc antibodyMyc Epitope Tag antibodyMyc protein antibodyMyc proto oncogene protein antibodyMyc proto-oncogene protein antibodyMyc-related translation/localization regulatory factor antibodyMYC_HUMAN antibodyMyc2 antibodymyca antibodyMyelocytomatosis oncogene a antibodyMyelocytomatosis oncogene antibodyNiard antibodyNird antibodyOncogene Myc antibodyOTTHUMP00000158589 antibodyOTTHUMP00000227763 antibodyProto-oncogene c-Myc antibodyProtooncogene homologous to myelocytomatosis virus antibodyRNCMYC antibodyTranscription factor p64 antibodyTranscriptional regulator Myc-A antibodyV-Myc avian myelocytomatosis viral oncogene homolog antibodyv-myc myelocytomatosis viral oncogene homolog (avian) antibodyzc-myc antibody

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

• 

  Western blot - anti-c-Myc antibody [9E10] (ab32)
  Anti-c-Myc antibody [9E10] - 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
• 

  Flow Cytometry-Anti-c-Myc antibody [9E10] - ChIP Grade(ab32)
  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.
  
  
• 

  Western blot - c-Myc antibody [9E10] (ab32)
  
  
  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.

• 

  Immunocytochemistry/ Immunofluorescence - c-Myc antibody [9E10] (ab32)
  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).
• 

  Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [9E10] - ChIP Grade (ab32)This image is courtesy of an anonymous Abreview
  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

• Western Blot