Overview

  • Product nameAnti-c-Myc antibody [Y69]
    See all c-Myc primary antibodies
  • Description
    Rabbit monoclonal [Y69] to c-Myc
  • SpecificityThis antibody is specific for c-Myc.
  • Tested applicationsWB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues in the N terminus of Human c-Myc.

  • Positive control
  • General notes

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    This product is available conjugated to DyLight® 488 or DyLight® 594 see ab139907 and ab139940 respectively.

    This product is available conjugated to Agarose valided in IP usage - ab178457

    A 40 µl trial size is available to purchase for this product.

    Myc is involved in MAPK-p38 signaling pathway -  see the interactive version.

Properties

Applications

Our Abpromise guarantee covers the use of ab32072 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
WB
IHC-P
ICC/IF
IP
  • Application notesICC/IF: 1/250 - 1/500.
    IHC-P: 1/50.
    IP: 1/150.
    WB: 1/10000. Detects a band of approximately 57 kDa (predicted molecular weight: 49 kDa).

    Is unsuitable for FACS.

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • 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
      • Cellular myelocytomatosis oncogene antibody
      • Class E basic helix-loop-helix protein 39 antibody
      • MRTL antibody
      • Myc antibody
      • Myc protein antibody
      • Myc proto oncogene 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
      • V-Myc avian myelocytomatosis viral oncogene homolog antibody
      • v-myc myelocytomatosis viral oncogene homolog (avian) antibody
      see all

    Anti-c-Myc antibody [Y69] images

    • ab32072 showing positive staining in Colonic adenocarcinoma tissue.

    • ab32072 showing positive staining in Lung adenocarcinoma tissue.

    • ab32072 staining c-Myc in HEK-293 cells transfected with CACNB4-c-Myc by Immunocytochemistry/ Immunofluorescence.
      Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100 then blocked using 5% serum for 20 minutes at 25°C. Samples were then incubated with ab32072 at a 1/250 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal, used at a 1/500 dilution.

      See Abreview

    • All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/1000 dilution

      Lane 1 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate
      Lane 2 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
      Lane 3 : THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate
      Lane 4 : A20 (Mouse B lymphoma cell line) Whole Cell Lysate
      Lane 5 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
      developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 49 kDa
      Observed band size : 57 kDa (why is the actual band size different from the predicted?)

      The predicted molecular weight of c-Myc is 48 kDa (SwissProt), however we expect to observe a banding pattern at 57 kDa.

      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab32072 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

    • c-Myc was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit monoclonal to c-Myc [Y69] 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, Jurkat whole cell extract 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 ab32072.

      Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

      Band: 57kDa; c-Myc [Y69]

    • Ab32072, at a dilution of 1/250, staining Hela cells by Immunoflourescence.
    • ab32072 showing positive staining in Gastric adenocarcinoma tissue.

    • ab32072 showing positive staining in Urinary bladder transitional carcinoma tissue.

    • Ab32072, at a dilution of 1/50, staining c-Myc in paraffin embedded human skin carcinoma tissue by Immunohistochemistry.
    • ICC/IF image of ab32072 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32072, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Anti-c-Myc antibody [Y69] (ab32072) at 1/10000 dilution + Jurkat cell lysate.

      Predicted band size : 49 kDa
      Observed band size : 57 kDa (why is the actual band size different from the predicted?)
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References for Anti-c-Myc antibody [Y69] (ab32072)

    This product has been referenced in:

    See all 41 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (SK-N-AS)
    Loading amount 30 µg
    Specification SK-N-AS
    Gel Running Conditions Reduced Denaturing (10% SDS gel)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted May 03 2013

    Hello and thank you for contacting us. The antibody ab32072 would be to detect the c-myc protein. Please let me know if you have any further questions.

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM Citrate
    Sample Human Tissue sections (Colon)
    Specification Colon
    Permeabilization No
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted May 30 2014

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step None
    Sample Human Tissue sections (UBC)
    Specification UBC
    Permeabilization No
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Apr 02 2014

    Application Western blot
    Loading amount 15 µg
    Gel Running Conditions Reduced Denaturing (any kDa gel)
    Sample Human Cell lysate - whole cell (A431 cells)
    Specification A431 cells
    Treatment EGF for 24 hours
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Username

    Felicia Cooper

    Verified customer

    Submitted Mar 19 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Mouse Tissue sections (Pancreas)
    Specification Pancreas
    Fixative Formaldehyde
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate pH 6
    Permeabilization No
    Blocking step 1% BSA 1% FBS as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Mar 13 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (SW480 cells, HCT116 cells)
    Loading amount 100000 cells
    Specification SW480 cells, HCT116 cells
    Gel Running Conditions Reduced Denaturing (4-12%)
    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 Dec 17 2012

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (4-20%)
    Sample Human Cell lysate - whole cell (HEK293 cells)
    Specification HEK293 cells
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Dr. Xiuquan Luo

    Verified customer

    Submitted Aug 13 2014

    Application Western blot
    Loading amount 100000 cells
    Gel Running Conditions Reduced Denaturing (10% gel)
    Sample Human Cell lysate - whole cell (MCF-7)
    Specification MCF-7
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Aug 12 2014

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step None
    Sample Human Tissue sections (tooth)
    Specification tooth
    Permeabilization No
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Apr 04 2014

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