Recombinant
RabMAb

Anti-c-Myc (phospho S62) antibody [EPR17924] (ab185656)

Overview

  • Product name
    Anti-c-Myc (phospho S62) antibody [EPR17924]
    See all c-Myc primary antibodies
  • Description
    Rabbit monoclonal [EPR17924] to c-Myc (phospho S62)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Dot blot, IHC-P, ICC/IF, IP, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human c-Myc aa 50-150 (phospho S62). The exact sequence is proprietary.
    Database link: P01106

  • Positive control
    • WB: HeLa, NIH/3T3 and C6 whole cell lysates. IHC-P: Human endometrium cancer, mouse spleen and rat testis tissues. ICC/IF: HeLa cells. IP: HeLa whole cell lysate treated with 200nM TPA for 10 minutes. FC: HeLa cells.
  • General notes

     

     

    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 ab185656 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
Dot blot 1/1000.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
IP 1/50.
WB 1/2000. Detects a band of approximately 57 kDa (predicted molecular weight: 48 kDa).
Flow Cyt 1/150.

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

 

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

  • Immunohistochemical analysis of paraffin-embedded Human endometrium cancer tissue labeling c-Myc (phospho S62) with ab185656 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on cancer cells of Human endometrial cancer is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling c-Myc (phospho S62) with ab185656 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing nuclear staining on HeLa cells.
    The staining decreased after blocking with phospho peptide (100μg/ml) overnight.

    The control peptide is a non-phospho peptide.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab185656 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling c-Myc (phospho S62) with ab185656 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on rat testis is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • All lanes : Anti-c-Myc (phospho S62) antibody [EPR17924] (ab185656) at 1/2000 dilution

    Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
    Lane 2 : C6 (Rat glial tumor cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 48 kDa
    Observed band size: 57 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling c-Myc (phospho S62) with purified ab185656 at 1/150 dilution (red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

  • All lanes : Anti-c-Myc (phospho S62) antibody [EPR17924] (ab185656) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) treated with Lambda Phosphatase whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 48 kDa
    Observed band size: 57 kDa (why is the actual band size different from the predicted?)


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-c-Myc (phospho S62) antibody [EPR17924] (ab185656) at 1/5000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated with 200nM Calyculin A and 1uM Okadaic Acid for 60 minutes.

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 48 kDa
    Observed band size: 57 kDa (why is the actual band size different from the predicted?)


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling c-Myc (phospho S62) with ab185656 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear and cytoplasmic staining on mouse spleen is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • c-Myc (phospho S62) was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated with 200nM TPA for 10 minutes with ab185656 at 1/50 dilution.

    Western blot was performed from the immunoprecipitate using ab185656 at 1/1000 dilution.

    VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/1500 dilution.

    Lane 1: HeLa whole cell lysate treated with 200nM TPA for 10 minutes,10 µg (Input).

    Lane 2: ab185656 IP in HeLa whole cell lysate treated with 200nM TPA for 10 minutes.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185656 in HeLa whole cell lysate treated with 200nM TPA for 10 minutes.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • Dot blot analysis of c -Myc (phospho T58) peptide (Lane 1), c-Myc non-phospho peptide (a control peptide for c-Myc phospho T58) (Lane 2), c-Myc (phospho S62) peptide (Lane 3), and c-Myc non-phospho peptide (a control peptide for c-Myc phospho S62) (Lane 4), labeled using ab185656 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Lanes 1, 2 and 4 are control peptides, lane 3 contains the immunogen peptide.

    Exposure time=3 minutes.

References

This product has been referenced in:
  • Malchenko S  et al. Stabilization of HIF-1a and HIF-2a, up-regulation of MYCC and accumulation of stabilized p53 constitute hallmarks of CNS-PNET animal model. PLoS One 12:e0173106 (2017). WB ; Human . Read more (PubMed: 28249000) »
  • Janghorban M  et al. The tumor suppressor phosphatase PP2A-B56a regulates stemness and promotes the initiation of malignancies in a novel murine model. PLoS One 12:e0188910 (2017). Read more (PubMed: 29190822) »

See all 3 Publications for this product

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

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