SELENOPROTEIN STRUCTURE AND FUNCTION

Detalles Bibliográficos
Clasificación:Libro Electrónico
Formato: Electrónico eBook
Idioma:Inglés
Publicado: [S.l.] : ELSEVIER ACADEMIC PRESS, 2022.
Colección:Methods in enzymology ; 662.
Temas:
Selenoproteins.
Selenoproteins
S�el�enoprot�eines.
Acceso en línea:Texto completo
Tabla de Contenidos:
  • Intro
  • Selenoprotein Structure and Function
  • Copyright
  • Contents
  • Contributors
  • Preface
  • Chapter One: Examining xCT-mediated selenium uptake and selenoprotein production capacity in cells
  • 1. Introduction
  • 2. Measurement of selenium uptake and selenoprotein production as mediated by xCT transporter
  • 2.1. Evaluation of xCT expression of SLC7A11 and SLC3A2
  • 2.1.1. Technical tips for the non-heat-denaturing Western blot of xCT
  • 2.2. Measurement of extracellular thiols as readout of xCT function leading to uptake
  • 2.2.1. Protocol for Ellman�s test
  • 2.2.1.1. Ellman�s test for measuring xCT activity on extracellular reduction
  • 2.2.1.2. Materials and equipment
  • 2.2.1.3. Step-by-step method details
  • 2.2.1.4. Expected outcomes
  • 2.2.1.5. Quantification and statistical analysis
  • 2.3. Measurement of selenium uptake
  • 2.3.1. Protocol for fluorometric assay
  • 2.3.1.1. Measurement of cellular selenium uptake capacity using fluorometric assay
  • 2.3.1.2. Materials and equipment
  • 2.3.1.3. Step-by-step method details
  • 2.3.1.4. Expected outcomes
  • 2.3.1.5. Quantification and statistical analysis
  • 2.3.1.6. Advantages
  • 2.3.1.7. Safety considerations and standards
  • 2.3.2. Protocol for sample prep for ICP-MS analysis
  • 2.3.2.1. Measurement of cellular selenium uptake capacity using ICP-MS
  • 2.3.2.2. Materials and equipment
  • 2.3.2.3. Step-by-step method details
  • 2.3.2.4. Example condition for ICP-MS run
  • 2.3.2.5. Expected outcomes
  • 2.3.2.6. Quantification and statistical analysis
  • 2.3.2.7. Advantages
  • 2.3.2.8. Limitations
  • 2.3.2.9. Safety considerations and standards
  • 2.4. Western blotting of selenoproteins as final readout
  • 2.4.1. Technical tips for the Western blotting of selenoproteins
  • 3. Summary
  • Acknowledgment
  • References
  • Chapter Two: Methods for accurate and reproducible studies of pharmacological effects of selenium in cancer
  • 1. Introduction
  • 2. Selenium cytotoxicity in cancer cell lines
  • 2.1. Effects of cell culture media composition on IC50 determinations of selenium compounds in cancer cell lines
  • 2.2. Phenotype expression depending on cell culture media
  • 2.3. Protocol for adherent cells
  • 2.3.1. Materials
  • 2.3.2. Procedure
  • 2.4. Protocol for cells in suspension
  • 2.4.1. Preparation of cell blocks for suspension cells
  • 2.4.2. Materials and equipment
  • 2.4.3. Procedure
  • 2.5. The activity of the selenoprotein thioredoxin reductase (TXNRD) in cells cultured in different media
  • 2.6. The extracellular redox milieu
  • 2.7. Thiol measurements
  • 2.7.1. Materials
  • 2.7.2. Procedure
  • 2.7.3. Quantification
  • 2.8. Alternative method-Thiol measurement by DTDP
  • 2.8.1. Buffers and chemical preparation
  • 2.8.2. Materials and equipment
  • 2.8.3. Procedure
  • 2.8.4. Expected outcomes
  • 2.8.5. Quantification
  • 2.8.6. Advantage

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