Recombinant
RabMAb

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

Overview

  • Product name
    Anti-c-Myc antibody [Y69]
    See all c-Myc primary antibodies
  • Description
    Rabbit monoclonal [Y69] to c-Myc
  • Specificity
    This antibody is specific for endogenous c-Myc.
  • Tested applications
    Suitable for: WB, ICC/IF, Flow Cyt, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human c-Myc aa 1-100 (N terminal). The exact sequence is proprietary.
    (Peptide available as ab166837)

  • Positive control
    • Purchase matching WB positive control:Recombinant human c-Myc protein
    • WB: Jurkat, Raji, MCF-7, K562, THP1, A20, rat spleen, L6, Neuro-2a and Raw264.7 cell lysates. ICC/IF: HeLa cells. IHC-P: Human skin carcinoma, diffuse large B cell lymphoma, adenocarcinoma of the colon, lung adenocarcinoma, gastric adenocarcinoma, urinary bladder transitional carcinoma tissues and esophagus. IP: Jurkat cell lysate. Flow Cyt: HeLa cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

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

    If you need conjugated anti-c-myc (Y69) RabMAb antibodies, find our range of products here.

    We also offer a PBS only version of this clone as product ab168727.

    For more information on choosing the right c-Myc antibody for you, please visit the following webpage: http://www.abcam.com/primary-antibodies/antibodies-to-c-myc-and-myc-tag 

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

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 1/10000. Detects a band of approximately 57 kDa (predicted molecular weight: 49 kDa).Can be blocked with c-Myc peptide (ab166837).
ICC/IF Use a concentration of 10 µg/ml.

1/100.

Flow Cyt 1/76.
IHC-P Use a concentration of 5 µg/ml.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP Use a concentration of 5 µg/ml.

Target

  • Function
    Participates 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 disease
    Note=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 similarities
    Contains 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 localization
    Nucleus > nucleoplasm. Nucleus > nucleolus.
  • Information by UniProt
  • Database links
  • Form
    c-Myc is also expressed in the cytoplasm.
  • Alternative names
    • AU016757 antibody
    • Avian myelocytomatosis viral oncogene homolog antibody
    • bHLHe39 antibody
    • c Myc 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-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

Images

  • IHC image of ab32072 staining c-Myc in human esophagus formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32072, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the Secondary only control (shown on the inset).

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab32072 staining c-Myc in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab32072 at 10μg/ml dilution (shown in green) and ab195889, mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling c-Myc with ab32072 at 1/500 dilution,followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.Nuclear staining on mouse spleen.Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • IHC image of ab32072 staining c-Myc in human adenocarcinoma formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32072, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Overlay histogram showing HeLa cells stained with ab32072 (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 (ab32072, 1/76 dilution) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) preadsorbed (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG [EPR25A] (monoclonal) (ab172730, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 nm bandpass filter. 

  • Immunocytochemistry/immunofluorescence analysis of HeLa cells labelling c-Myc with purified ab32072 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

  • Unpurified ab32072 staining c-Myc in HEK293 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

  • ICC/IF image of unpurified 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 anti rabbit DyLight® 488 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.

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

    Lane 1 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates
    Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
    Lane 4 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
    Lane 5 : THP-1 (Human monocytic leukemia monocyte) whole cell lysates
    Lane 6 : Rat spleen whole cell lysates
    Lane 7 : L6 (Rat skeletal muscle myoblast) whole cell lysates
    Lane 8 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
    Lane 9 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/20000 dilution

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


    Exposure time : 3 minutes

    Blocking and dilution buffer: 5% NFDM/TBST.

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

    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 HRP antibody, and visualised using ECL development solution ab133406.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human diffuse large B cell lymphoma tissue labelling c-Myc with purified ab32072 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of the colon tissue labelling c-Myc with purified ab32072 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skin carcinoma tissue labelling c-Myc with unpurified ab32072 at 1/50.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarinoma of colon tissue labelling c-Myc with unpurified ab32072.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung adenocarinoma tissue labelling c-Myc with unpurified ab32072.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric adenocarcinoma tissue labelling c-Myc with unpurified ab32072.

  • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human urinary bladder transitional carcinoma tissue labelling c-Myc with unpurified ab32072.

  • c-Myc was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of unpurified 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 unpurified ab32072.

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

    Band: 57kDa; c-Myc [Y69]

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Weng S  et al. Restoration of MYC-repressed targets mediates the negative effects of GM-CSF on RUNX1-ETO leukemogenicity. Leukemia 31:159-169 (2017). WB . Read more (PubMed: 27389055) »
  • Lu T  et al. Adamts18 deficiency promotes colon carcinogenesis by enhancing ß-catenin and p38MAPK/ERK1/2 signaling in the mouse model of AOM/DSS-induced colitis-associated colorectal cancer. Oncotarget 8:18979-18990 (2017). WB, IHC ; Mouse . Read more (PubMed: 28145888) »

See all 192 Publications for this product

Customer reviews and Q&As

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

The antibody ab32072 would be to detect the c-myc protein.

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

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colorectal liver metastasis)
Antigen retrieval step
Heat mediated
Permeabilization
Yes - TBS with 0.25% Triton for 10 min
Specification
Colorectal liver metastasis
Blocking step
Power Block Universal Blocking Reagent 10x as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Formaldehyde
Username

Mr. Michael Wuehrl

Verified customer

Submitted Jun 30 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Dog Tissue sections (Colon)
Permeabilization
Yes - Triton
Specification
Colon
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Fixative
Acetone
Username

Abcam user community

Verified customer

Submitted Apr 03 2017

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 (fetal liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: high pH CC1 (Roche Ventana)
Specification
fetal liver
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: 24°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 16 2017

1-10 of 40 Abreviews or Q&A

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

Sign up