Apoptosis a practical approach 1st Edition by GP Studzinski, BD Hames ISBN 9780191566004 0191566004

Apoptosis a practical approach 1st Edition by GP Studzinski, BD Hames – Ebook PDF Instant Download/Delivery. 9780191566004 ,0191566004
Full download Apoptosis a practical approach 1st Edition after payment

Product details:
ISBN 10: 0191566004
ISBN 13: 9780191566004
Author: GP Studzinski, BD Hames

Apoptosis a practical approach 1st Edition Table of contents:

1. Overview of apoptosis

1. General introduction and overview of contents

2. Historical perspective

3. Distinction of apoptosis from other forms of cell death

4. Apoptotic cascades

5. Time course of apoptotic cascades

6. Selection of methods

Procedure for determination of increased mitochondrial to nuclear DNA ratio, for the detection and q

7. Pitfalls

References

2. Morphological recognition of apoptotic cells

1. Introduction

Key morphological features of apoptotic cells

2. Light and fluorescent microscopy techniques for the assessment of apoptosis

Preparation of cell or tissue samples

Nuclear counterstains

3. Electron microscopic techniques

4. Quantitation of apoptotic events

Methods

Problems in scoring apoptotic events

5. In situ detection of DNA strand breaks

6. Other techniques

7. Conclusions

References

3. Assessment of DNA damage in apoptosis

1. Introduction

2. Types of DNA fragmentation

3. Measurement of high molecular weight DNA fragmentation by pulse-field gel electrophoresis

Pulse-field gel electrophoresis procedures

Strategies for electrophoretic separations

4. Detection of internucleosomal DNA fragmentation by standard agarose gel electrophoresis

5. Filter elution assay to measure apoptotic DNA fragmentation

Equipment

Filters and solutions

DNA labelling and preparation of experimental cell cultures

Protocols

Counting samples and computations

References

4. Analysis of cell death by flow and laser-scanning cytometry

1. Introduction

2. Preparation of cells for analysis by LSC. Detection of apoptotic cells based on changes in nuclea

3. Light-scattering properties of apoptotic and necrotic cells

4. Analysis of mitochondrial transmembrane potential (Δψ[sub(m)])

5. Detection of apoptotic cells exposing phosphatidylserine on plasma membrane

6. Detection of apoptotic cells based on their fractional DNA content

7. Identification of apoptotic cells based on the presence of DNA strand breaks

8. Identification of apoptotic cells based on the increased DNA sensitivity to denaturation

9. General comments

References

5. Cell-mediated cytotoxicity and cell death receptors

1. Overview of pathways for cell-mediated cytotoxicity

Fas-based cytotoxicity

Perforin/granzyme-based cytotoxicity

2. Standard methods for the evaluation of cell-mediated cytotoxicity

Chromium release assay

The JAM test

BLT–esterase release assay

Estimation of target cell nuclear fragmentation using fluorescent dyes

Activation-induced cell death (AICD)

3. Separate studies of Fas- and perforin/granzyme-based cytotoxicity

Fas-based cytotoxicity in the absence of perforin/granzyme contribution

Perforin/granzyme-based cytotoxicity in the absence of FasL contribution

4. Use of caspase and granzyme inhibitors in cell-mediated cytotoxicity assays

Caspase inhibitors

Granzyme inhibitors

References

6. Sphingolipids as messengers of cell death

1. Introduction

2. Sphingolipids and their role in apoptosis

Overview of Sphingolipids

Role of Sphingolipids in apoptosis

Strategies and considerations in evaluating Sphingolipids

3. Extraction of Sphingolipids and normalization by lipid phosphate

Extraction of sphingosines, ceramides, and sphingomyelins

Alkaline hydrolysis of Sphingolipids

Lipid phosphate measurement for normalization of experimental samples

4. Analysis of sphingoid backbones

HPLC on sphingoid backbones

5. Analysis of ceramides

6. Analysis of sphingomyelins

7. Measurement of SMase and SM synthase activities

References

7. Cytochemical detection of cytoskeletal and nucleoskeletal changes during apoptosis

1. Introduction

2. Cyto- and nucleoskeletal changes during apoptosis

General

Microfilaments and microfilament-associated proteins

Microtubules

Intermediate filaments

Nucleoskeletal components

Generation of neo-epitopes in cytoskeletal proteins during apoptosis

3. Apoptosis detection systems

Overview

The annexin V affinity assay

Detection of apoptosis using the TUNEL assay

References

8. Metabolic alterations associated with apoptosis

1. Introduction

2. Detection of changes in the mitochondrial transmembrane potential

3. Detection of intracellular reactive oxygen intermediates (ROI)

4. Determination of glutathione levels and its oxidative state

5. Measurement of catalase in cells undergoing apoptosis

References

9. Methods of measuring Bcl-2 family proteins and their functions

1. Introduction

2. Immunoblot analysis

Cell lysis and tissue processing procedures

Immunoblotting procedure

3. Detection of Bcl-2 family proteins by flow cytometry (FACS)

Indirect immunofluorescence procedure

Antibodies

4. Northern blot analysis of BCL-2 mRNA

General guidelines for RNA blotting

Pre-hybridization and hybridization procedures

Solutions

5. Reverse transcriptase-polymerase chain reaction (RT-PCR)

cDNA synthesis

PCR amplication

Southern blot analysis of PCR products

Primers and probes for PCR

6. Immunohistochemical detection of Bcl-2 family proteins in tissues

Procedures for tissue preparation, embedding, and sectioning

Pre-staining sample preparation

Immunostaining

Counterstaining and mounting

Solutions

7. Immunohistochemical detection of Bcl-2 protein in cultured cells

Procedures

Immunostaining

Solutions

8. Production of recombinant Bcl-2 family proteins in bacteria

Human Bcl-2

Human Bcl-X[sub(L)] protein

Mouse Bax protein

Human Bid protein

9. Measurements of pore formation by Bcl-2 family proteins

Liposome preparation

Channel activity measurements

10. Methods of assaying Bcl-2 and Bax family protein function in yeast

Plasmid considerations

Saccharomyces cerevisiae strains

Media and growth conditions

Transformation of S. cerevisiae by the LiOAc method

Assay for Bax-induced lethality in S. cerevisiae

11. Summary

References

10. Methods for detecting proteolysis during apoptosis in intact cells

1. Introduction

2. Involvement of proteases in apoptosis

3. Caspase nomenclature and classification

4. Detection of procaspases and their cleavage products by immunoblotting

Theoretical considerations

Practical considerations in immunoblotting for caspases

Sample preparation and immunoblotting

5. Cleavage of caspase targets

Establishing that proteases are activated in situ

Establishing that a polypeptide is cleaved by caspases

6. Assays of caspase activity

Theoretical considerations of caspase activity assays

Measuring caspase activity in cell lysates

7. Detection of active caspases by affinity labelling

Theoretical considerations in affinity labelling

Practical considerations in the use of affinity labelling reagents

Labelling and detecting enzymatically active caspases

8. Studying the biological effects of caspase cleavages

A molecular approach for analysis of the effects of substrate cleavage

Use of caspase inhibitors: promises and pitfalls

References

List of Suppliers

Index

People also search for Apoptosis a practical approach 1st Edition:

apoptosis assay

apoptosis experiment

a topical approach to lifespan development 11th edition access code

apoptosis ap biology