Overview

Description

  • Nature
    Recombinant
  • Source
    Escherichia coli
  • Amino Acid Sequence
    • Accession
    • Species
      Human
    • Sequence
      MDFFRVVENQQPPATMPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQ QQSELQPPAPSEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGD NDGGGGSFSTADQLEMVTELLGGDMVNQSFICDPDDETFIKNIIIQDCMW SGFSAAAKLVSEKLASYQAARKDSGSPNPARGHSVCSTSSLYLQDLSAAA SECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDSLLSSTESSPQGSP EPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGSPSAG GHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQ ISNNRKCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELE NNEKAPKVVILKKATAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLR NSCAESGGGGSPGRRRRRRRRRRR
    • Molecular weight
      53 kDa
    • Amino acids
      1 to 454
    • Additional sequence information
      Please note that ab169901 is isoform 2 of UniProt accession P01106.

Specifications

Our Abpromise guarantee covers the use of ab169901 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Biological activity

    Activity: Reprogramming mouse fibroblast cell to iPS cells using 3 retroviral vectors, which carry Oct4, Sox2 & Klf4 with this protein as replacement assay. 8 μg/ml of human Klf4-11R were added in reprogramming medium every 48 hours for 20 days. Intracellular protein penetration rate was tested using DyLight labeled ab169901 protein at 1 μg/ ml for 30 min incubation for human fibroblast cells at 37oC. More than 90% cell will be positive one hour after sample incubation.

    Cellular Toxicity: This recombinant protein was tested on mouse embryonic stem cells up to 50 µg/ml in culture medium. Suggested reprogramming protein concentration is between 0.5 to 8 μg/ml for both human and mouse fibroblast cells applications.

  • Applications

    Western blot

    Functional Studies

    SDS-PAGE

    Mass Spectrometry

  • Purity
    >90% by SDS-PAGE.
    ab169901 was expressed in E. coli as inclusion bodies, solubilized, refolded, and further purified.
  • Form
    Liquid
  • Additional notes
    ab169901 is fused to an eleven arginine (11R) membrane penetration domain at the C terminus to enable penetratation across the plasma membrane of mammalian cells.
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

    pH: 7.50
    Constituent: 0.24% Tris
    Note: Contains NaCl , KCl, CaCl2, MgCl2, arginine, DTT, and glycerol.

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

General Info

  • Alternative names
    • AU016757
    • Avian myelocytomatosis viral oncogene homolog
    • bHLHe39
    • c Myc
    • Class E basic helix-loop-helix protein 39
    • MRTL
    • Myc
    • Myc protein
    • Myc proto oncogene protein
    • Myc proto-oncogene protein
    • myc-related translation/localization regulatory factor
    • MYC_HUMAN
    • Myc2
    • MYCC
    • Myelocytomatosis oncogene
    • Niard
    • Nird
    • Oncogene Myc
    • OTTHUMP00000158589
    • Proto-oncogene c-Myc
    • Protooncogene homologous to myelocytomatosis virus
    • RNCMYC
    • Transcription factor p64
    • Transcriptional regulator Myc-A
    • V-Myc avian myelocytomatosis viral oncogene homolog
    • v-myc myelocytomatosis viral oncogene homolog (avian)
    see all
  • 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

References

This product has been referenced in:
  • Tarrado-Castellarnau M  et al. De novo MYC addiction as an adaptive response of cancer cells to CDK4/6 inhibition. Mol Syst Biol 13:940 (2017). Read more (PubMed: 28978620) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Functional Studies
Kinase assay.
Recombinant human c-MYC protein (Abcam, ab169901) was prepared along with required cofactors in freshly prepared Base Reaction Buffer (20 mM Hepes pH 7.5, 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na3VO4, 2 mM DTT, 1% DMSO). CDK4-Cyclin D1 or CDK6-Cyclin D1 complexes were delivered into the substrate solution at varying concentrations and gently mixed. A kinase titration without substrate was also performed to determine the background signal, and Retinoblastoma protein was used as a substrate to obtain a positive phosphorylated control. 33P-ATP was added into the reaction mixture to initiate the reaction and incubated for 60 and 120 min at room temperature. Reactions were spotted onto P81 ion exchange filter paper. Unbound phosphate was removed by washing filters extensively in 0.75% phosphoric acid. 33P signal was determined using Typhoon phosphorimagers. For mass spectrometry phosphorylation site profiling and western blot, cold ATP was used instead of 33P-ATP.
We detected specific 33P signals in both kinase reactions, indicating that both CDK4-Cyclin D1 and CDK6-Cyclin D1 complexes directly phosphorylate MYC.
By analyzing MYC tryptic peptides by mass spectrometry after the kinase assay, we found that peptides KFELLPT(phosphor)PPLSPSR and KFELLPTPPLS(phosphor)PSRR were phosphorylated on Threonine 7 (corresponding to c-MYC T58) and Serine 11 (corresponding to c-MYC S62), respectively.
Detection of P-MYC (Ser62) by Western blot. Western blot analysis after anti-P-MYC (Ser62) (ab106952) antibody incubation showed detection of P-MYC (Ser62) in samples that were incubated in presence of ATP during the kinase assay.

Abcam user community

Verified customer

Submitted Sep 25 2018

Answer

The lab has indicated they do not purify using a tag, but that they use multiple steps including intensive purification of inclusion bodies and size exclusion chromatography. They do not add tags because they want to avoid their potential effects to protein's activities.

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