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标题:Methods for Assessing Apoptosis and Anoikis in Normal Intestine/Colon and Colorectal Cancer
时间:2019-11-08 23:00:12
DOI:10.1007/978-1-4939-7765-9_7
大小:1044 kb
页数:40 PAGES
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目录:
  • Chapter 7: Methods for Assessing Apoptosis and Anoikis in Normal Intestine/Colon and Colorectal Cancer
    • 1 Introduction
    • 2 Materials
      • 2.1 Cell Lines
        • 2.1.1 Cell Culture
      • 2.2 Tissues
        • 2.2.1 Cryo-Embedding for Cryostat Sectioning (See Note 5)
        • 2.2.2 Ex Vivo Explant Culture (See Note 6)
      • 2.3 Apoptosis Induction
      • 2.4 Anoikis Induction
      • 2.5 Apoptosis/Anoikis Assays
        • 2.5.1 ISEL (In Situ TUNEL [Terminal Deoxynucleotidyl Transferase (TdT) dUTP-Mediated Nick-End Labeling])
        • 2.5.2 Annexin V Staining
        • 2.5.3 DNA Laddering Assays
        • 2.5.4 SDS-PAGE and Western Blotting
        • 2.5.5 Fluorogenic Caspase Activity Assays
    • 3 Methods
      • 3.1 ISEL Detection of Apoptosis
        • 3.1.1 Twenty-Week-Old Human Fetal Small Intestine (See Note 29)
        • 3.1.2 18 Weeks-Old Human Fetal Colon Explants
        • 3.1.3 Cell Cultures
      • 3.2 Detection of Apoptosis-Associated Phosphatidylserine (PS) Translocation to Cell Surface
        • 3.2.1 Cell Cultures
      • 3.3 Visualization of Caspase-Dependent Internucleosomal DNA Fragmentation (``DNA Laddering´´)
        • 3.3.1 Apoptosis Induction
        • 3.3.2 Anoikis Induction
      • 3.4 Assessment of CASP-3 Activation/Activity by Western Blotting
        • 3.4.1 Apoptosis, Anoikis Induction
        • 3.4.2 SDS-PAGE and Western Blotting
      • 3.5 Direct Assessment of Caspase Activity during Apoptosis, Anoikis
        • 3.5.1 Apoptosis Induction
        • 3.5.2 Time-Course Kinetic of CASP-8, CASP-9 and CASP-3 Activity Following Anoikis Induction
        • 3.5.3 Time-Course Kinetic of CASP-3 Activity for Evidence of Anoikis Resistance Displayed by CRC Cells
    • 4 Notes
    • References

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