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标题:Expression, purification and functional characterization of Wnt signaling co-receptors LRP5 and LRP6.
时间:2019-09-12 00:11:06
DOI:10.1016/j.pep.2009.08.012
PMID:19716419
作者:Fleury D;Vayssiere B;Touitou R;Gillard C;Lebhar H
出版源: 《Protein Expression & Purification》 ,2010 ,70 (1) :39-47
摘要:C, Lebhar H, Rawadi G, and Mollat P (2010) Expression, purification and functional characterization of Wnt signaling co-receptors LRP5 and LRP6. Protein...
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页数:10 PAGES
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目录:
  • Expression, purification and functional characterization of Wnt signaling co-receptors LRP5 and LRP6
    • Introduction
    • Materials and methods
      • Reagents
      • Cloning and expression of EC-LRP5 and EC-LRP6
      • Cloning and expression of mDkk1-HRP and mDkk2-HRP
      • Protein purification
        • Purification of EC-LRP6 from culture medium
        • Purification of EC-LRP5 from culture medium
        • mDkk1-HRP purification
        • mDkk2-HRP purification
      • Measurement of luminescence for the purification samples
      • Binding assays
        • Luminescent interaction assay (LIA)
        • Competition assay with mDkk1-V5-6His
      • Wnt signaling cell-based assay
      • Analysis on SDS–PAGE and western blot
    • Results
      • Expression of EC-LRP5 and 6
      • EC-LRP6 purification
      • EC-LRP5 purification
      • Expression of mDkk1 and mDkk2-HRP
      • mDkk1-HRP purification
      • mDkk2-HRP purification
      • Functional activity of purified proteins
      • Binding assays (LIA)
    • Discussion
    • Acknowledgments
    • References

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