Overview

  • Product nameAnti-c-Myc (phospho T58) antibody
    See all c-Myc primary antibodies
  • Description
    Rabbit polyclonal to c-Myc (phospho T58)
  • Tested applicationsSuitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Sheep, Chicken, Cat, Pig, Chimpanzee, Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human c-Myc.

  • Positive control
    • WB: HeLa and NIH3T3 whole cell lysates. ICC/IF: methanol fixed MCF7 cells.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab85380 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 Use a concentration of 1 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 49 kDa).
ICC/IF Use a concentration of 5 µg/ml.

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. Phosphorylation at Ser-329 by PIM2 leads to the stabilization of MYC (By similarity). Phosphorylation at Ser-62 by CDK2 prevents Ras-induced senescence.
    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
  • Alternative names
    • 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
    • Niard antibody
    • Nird antibody
    • Proto-oncogene c-Myc antibody
    • Transcription factor p64 antibody
    • v myc avian myelocytomatosis viral oncogene homolog antibody
    • v myc myelocytomatosis viral oncogene homolog antibody
    see all

Anti-c-Myc (phospho T58) antibody images

  • Lanes 1 - 4 : Anti-CA5B antibody (ab85280) at 1 µg/ml
    Lanes 5 - 6 : Anti-c-Myc (phospho T58) antibody (ab85380) at 1 µg/ml

    Lane 1 : NIH3T3 (-AP)
    Lane 2 : NIH3T3 (+AP)
    Lane 3 : NIH3T3 + C peptide with c-Myc peptide (ab116700)
    Lane 4 : NIH3T3 + P peptide with c-Myc (phospho T58) peptide (ab104386)
    Lane 5 : Raji (-AP)
    Lane 6 : Raji (+AP)

    Lysates/proteins at 20 µg per lane.


    Performed under reducing conditions.

    Predicted band size : 49 kDa

    The blots were produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membranes were blocked for an hour, membrane strips 2 and 6 were incubated with alkaline phospatase (100 U per mL) and strips 1 and 5 in phosphatase reaction buffer without enzyme for one hour at 37 degrees before incubating lane 1 - 4 with ab85380 (rabbit polyclonal to c-Myc (phospho T58) antibody; 1 ug per mL) and lane 5-6 with ab32072 (rabbit monoclonal  to c-Myc; 1:10000 dilution), in the presence of loading control ab8245 (mouse anti-GAPDH antibody; diluted 1:20000) for 24 hours at 4°C. Prior to adding the primary antibodies to the strips 3-4, 1 ug per mL of ab116700 (c-Myc control peptide: lane 3) or ab104386 (c-Myc (phospho T58) peptide: lane 4) were added to the antibodies.

  • Serially diluted ab85380 was bound to immobilised phospho (ab104386) - or control (ab116700) peptides (1 microgram x mL-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 50000 times) and signal was developed with TMB substrate.

  • All lanes : Anti-c-Myc (phospho T58) antibody (ab85380) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) with c-Myc (phospho T58) peptide (ab104386) at 1 µg/ml
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) with c-Myc peptide (ab116700) at 1 µg/ml
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 49 kDa
    Observed band size : 52 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 38 kDa,71 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 16 minutes
    c-Myc contains a potential glycosylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • Anti-c-Myc (phospho T58) antibody (ab85380) at 1 µg/ml + NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 49 kDa
    Observed band size : 55 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 40 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • ICC/IF image of ab85380 stained MCF-7 cells. The cells were 100% methanol fixed (5 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 ab85380 at 5µg/ml overnight at +4°C. The secondary antibody (green) was a goat anti-rabbit DyLight® 488(ab96899) IgG (H+L) used at a 1/1000 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.

References for Anti-c-Myc (phospho T58) antibody (ab85380)

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