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Recombinant human c-Myc protein (Active) (ab169901)

Reviews (3)Q&A (1)References (4)

Key features and details

  • Expression system: Escherichia coli
  • Purity: > 93% SDS-PAGE
  • Active: Yes
  • Suitable for: WB, Functional Studies, SDS-PAGE, MS

製品の詳細

  • 製品名

    Recombinant human c-Myc protein (Active)
    c-Myc タンパク質・ペプチド 製品一覧

  • 生理活性

    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.

  • 精製度

    > 93 % SDS-PAGE.
    ab169901 was expressed in E. coli as inclusion bodies, solubilized, refolded, and further purified.

  • エンドトキシン・レベル

    5.000 Eu/µg

  • 発現系

    Escherichia coli

  • アクセッション番号

    P01106-2

  • タンパク質長

    Full length protein

  • Animal free

    No

  • 由来

    Recombinant

    • 生物種

      Human

    • 配列

      MDFFRVVENQQPPATMPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQ QQSELQPPAPSEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGD NDGGGGSFSTADQLEMVTELLGGDMVNQSFICDPDDETFIKNIIIQDCMW SGFSAAAKLVSEKLASYQAARKDSGSPNPARGHSVCSTSSLYLQDLSAAA SECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDSLLSSTESSPQGSP EPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGSPSAG GHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQ ISNNRKCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELE NNEKAPKVVILKKATAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLR NSCAESGGGGSPGRRRRRRRRRRR

    • 予測される分子量

      53 kDa

    • 領域

      1 to 454

    • 配列の追加情報

      Please note that ab169901 is isoform 2 of UniProt accession P01106. (NP_002458.2) C-terminal 11R tag : ESGGGGSPGRRRRRRRRRRR

特性

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.

  • アプリケーション

    Western blot

    Functional Studies

    SDS-PAGE

    Mass Spectrometry

  • 製品の状態

    Liquid

  • 備考

    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.

    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 ug / ml for both human and mouse fibroblast cells applications.

     

  • Concentration information loading...

前処理および保存

  • 保存方法および安定性

    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

    Proprietary formulation of 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.

関連情報

  • 別名

    • AU016757
    • Avian myelocytomatosis viral oncogene homolog
    • bHLHe39
    • c Myc
    • Cellular myelocytomatosis oncogene
    • Class E basic helix-loop-helix protein 39
    • MGC105490
    • MRTL
    • Myc
    • Myc protein
    • Myc proto oncogene protein
    • Myc proto-oncogene protein
    • myc-related translation/localization regulatory factor
    • MYC_HUMAN
    • Myc2
    • myca
    • MYCC
    • Myelocytomatosis oncogene
    • Myelocytomatosis oncogene a
    • Niard
    • Nird
    • oncogene c-Myc
    • Oncogene Myc
    • OTTHUMP00000158589
    • OTTHUMP00000227763
    • 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)
    • zc-myc
    see all

  • 機能

    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.

  • 関連疾患

    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.

  • 配列類似性

    Contains 1 basic helix-loop-helix (bHLH) domain.

  • 翻訳後修飾

    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.

  • 細胞内局在

    Nucleus > nucleoplasm. Nucleus > nucleolus.
  • Information by UniProt

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献 (4)

ab169901 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab169901 は 4 報の論文で使用されています。

  • Nowak DG  et al. The PHLPP2 phosphatase is a druggable driver of prostate cancer progression. J Cell Biol 218:1943-1957 (2019). PubMed: 31092557
  • Blake DR  et al. Application of a MYC degradation screen identifies sensitivity to CDK9 inhibitors in KRAS-mutant pancreatic cancer. Sci Signal 12:N/A (2019). PubMed: 31311847
  • Han H  et al. Small-Molecule MYC Inhibitors Suppress Tumor Growth and Enhance Immunotherapy. Cancer Cell 36:483-497.e15 (2019). PubMed: 31679823
  • 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). PubMed: 28978620

レビューと Q&A

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1-4 of 4 Abreviews or Q&A

Direct ELISA with MYC protein

 Excellent
Abreviews
abreview image
Application
ELISA
MYC protein was coated onto a plate at a concentration of 1 nM. Plate was washed 3X with PBS-T. Anti-MYC primary antibody (ab32072) was titrated and incubated for 2 hr. Plate was washed 3X with PBS-T. Secondary antibody (HRP anti-rabbit) was incubated for 1 hr. ELISA was developed using TMB high sensitivity reagent.

Abcam user community

Verified customer

投稿 Nov 25 2020

Kinase assay - mass spectrometry identification of CDK9 phosphorylation of MYC

 Excellent
Abreviews
abreview image
Application
Functional Studies
Recombinant MYC (Abcam ab169901) at 1000 nM was incubated with 50 nM recombinant CDK9 together with cyclin T1 required for kinase activity (Thermo Fisher PV4131) and 100 μM ATP for 30 min at 30°C. The reaction was stopped by adding SDS sample buffer and boiling for 5 min at 95°C. After separation by SDS-PAGE, the gel was stained for 30 min with SimplyBlue Coomassie and destained overnight in water. Protein bands corresponding to MYC were excised and the proteins were reduced, alkylated, and in-gel digested with trypsin overnight at 37˚C. Peptides were extracted and dried by vacuum centrifugation. Samples were analyzed by LC/MS/MS using an Easy nLC 1000 coupled to a QExactive HF mass spectrometer.

Using a kinase assay followed by mass spectrometry, we identified putative CDK9 phosphorylation sites on MYC.

Devon Blake

Verified customer

投稿 Aug 10 2020

In vitro kinase assays with CDK4/6-Cyclin D1 complexes and full-lenght recombinant human c-MYC protein

 Excellent
Abreviews
abreview image
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

投稿 Sep 25 2018

Question

How was the protein ab169901 (with flexible linker domain and an eleven arginine tag) purified? We think that is necessary to add a tag as His or GST.

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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.

Read More

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