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重组人c-Myc蛋白(Active) (ab169901)

Reviews (3)Q&A (1)References (11)

Key features and details

  • Expression system: Escherichia coli
  • Purity: > 93% SDS-PAGE
  • Endotoxin level: = 5.000 Eu/µg
  • Active: Yes
  • Suitable for: WB, Functional Studies, SDS-PAGE, MS

描述

  • 产品名称

    重组人c-Myc蛋白(Active)
    参阅全部 c-Myc 蛋白酶

  • 生物活性

    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.

  • 纯度

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

  • 内毒素水平

    = 5.000 Eu/µg

  • 表达系统

    Escherichia coli

  • Accession

    P01106-2

  • 蛋白长度

    Full length protein

  • 无动物成分

    No

  • 性质

    Recombinant

    • 种属

      Human

    • 序列

      MDFFRVVENQQPPATMPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQ QQSELQPPAPSEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGD NDGGGGSFSTADQLEMVTELLGGDMVNQSFICDPDDETFIKNIIIQDCMW SGFSAAAKLVSEKLASYQAARKDSGSPNPARGHSVCSTSSLYLQDLSAAA SECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDSLLSSTESSPQGSP EPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGSPSAG GHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQ ISNNRKCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELE NNEKAPKVVILKKATAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLR NSCAESGGGGSPGRRRRRRRRRRR

    • 预测分子量

      53 kDa

    • 氨基酸

      1 to 454

    • 额外的序列信息

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

技术指标

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.

  • 应用

    Western blot

    Functional Studies

    SDS-PAGE

    Mass Spectrometry

  • 形式

    Liquid

  • 补充说明

    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...

制备和贮存

  • 稳定性和存储

    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.

常规信息

  • 别名

    • 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

  • 功能

    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.

  • 疾病相关

    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.

  • 序列相似性

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

  • 翻译后修饰

    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.

  • 细胞定位

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

实验方案

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

文献 (11)

发表研究结果有使用 ab169901？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab169901 被引用在 11 文献中.

  • Boulos JC  et al. In Silico, In Vitro, and In Vivo Investigations on Adapalene as Repurposed Third Generation Retinoid against Multiple Myeloma and Leukemia. Cancers (Basel) 15:N/A (2023). PubMed: 37627164
  • Elbadawi M  et al. The Novel Artemisinin Dimer Isoniazide ELI-XXIII-98-2 Induces c-MYC Inhibition, DNA Damage, and Autophagy in Leukemia Cells. Pharmaceutics 15:N/A (2023). PubMed: 37111592
  • Lepore Signorile M  et al. c-MYC Protein Stability Is Sustained by MAPKs in Colorectal Cancer. Cancers (Basel) 14:N/A (2022). PubMed: 36230763
  • Muto T  et al. TRAF6 functions as a tumor suppressor in myeloid malignancies by directly targeting MYC oncogenic activity. Cell Stem Cell 29:298-314.e9 (2022). PubMed: 35045331
  • Dahiya NR  et al. The Sin3A/MAD1 Complex, through Its PAH2 Domain, Acts as a Second Repressor of Retinoic Acid Receptor Beta Expression in Breast Cancer Cells. Cells 11:N/A (2022). PubMed: 35406744
View all Publications for this product

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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.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 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.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Devon Blake

Verified customer

提交于 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.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 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.

Read More
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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