Immunochemical Protocols

1. Front Matter

   Pages i-xiii

   PDF

2. Production of Polyclonal Antisera

   • Jonathan A. Green, Margaret M. Manson

   Pages 1-5

3. Efficient Elution of Purified Proteins from Polyvinylidene Difluoride Membranes (lmmobilon) After Transfer from SDS-PAGE and Their Use as Immunogens

   • Boguslaw Szewczyk, Donald F. Summers

   Pages 7-12

4. Raising Polyclonal Antibodies Using Nitrocellulose-Bound Antigen

   • Monique Diano, André Le Bivic, Michel Hirn

   Pages 13-22

5. Synthesis of Peptides for Use as Immunogens

   • David C. Hancock, Gerard I. Evan

   Pages 23-32

6. Production and Characterization of Antibodies Against Synthetic Peptides

   • David C. Hancock, Gerard I. Evan

   Pages 33-41

7. Preparation and Testing of Monoclonal Antibodies to Recombinant Proteins

   • Christopher J. Dean

   Pages 43-63

8. Screening of Monoclonal Antibodies Using Antigens Labeled with Acetylcholinesterase

   • Yveline Frobert, Jacques Grassi

   Pages 65-78

9. Purification of Immunoglobulin G (IgG)

   • Michael G. Baines, Robin Thorpe

   Pages 79-104

10. Epitope Mapping

    • Sara E. Mole

    Pages 105-116

11. Enzyme-Antienzyme Method for Immunohistochemistry

    • Michael G. Ormerod, Susanne F. Imrie

    Pages 117-124

12. double Label Immunohistochemistry on Tissue Sections Using Alkaline Phosphatase and PeroxIDase Conjugates

    • Jonathan A. Green, Margaret M. Manson

    Pages 125-129

13. Immunohistochemical detection of BromodeoxyurIDine-Labeled Nuclei for In Vivo Cell Kinetic Studies

    • Jonathan A. Green, Richard E. Edwards, Margaret M. Manson

    Pages 131-135

14. Avidin–Biotin Technology

    • Edward A. Bayer, Meir Wilchek

    Pages 137-142

15. Avidin–Biotin Technology

    • Edward A. Bayer, Meir Wilchek

    Pages 143-148

16. Immunochemical Applications of Avidin-Biotin Technology

    • Edward A. Bayer, Meir Wilchek

    Pages 149-162

17. Preparation of Gold Probes

    • Julian E. Beesley

    Pages 163-168

18. Immunogold Probes for Light Microscopy

    • Julian E. Beesley

    Pages 169-176

19. Immunogold Probes in Electron Microscopy

    • Julian E. Beesley

    Pages 177-185

20. Electron Microscopic Silver Enhancement for Double Labeling with Antibodies Raised in the Same Species

    • Kurt Bienz, Denise Egger

    Pages 187-193

Molecular biologists can no longer afford to be experts in only one par­ ticular area. They need to be equally competent at handling DNA, RNA, and proteins, moving from one area to another as required by the problem. Ad­ vances in the field have been greatly helped by numerous applications of antibody-antigen interactions, so that a basic knowledge of immunology has now become useful. The emphasis of Immunochemical Protocols is not on the production of antibodies, since many people will beg or buy these, but rather on the enor­ mous range of applications to which these powerful reagents can be put. For completeness, however, methods are included for the production of both polyclonal and monoclonal antibodies. Techniques that may be helpful in obtaining antibodies to elusive antigens and to synthetic peptides are also provided. Both qualitative and quantitative methods are described for local­ izing, characterizing, and quantifying various antigens. These range from the use of antibodies in screening DNA libraries to their application in EUSA assays for quantifying adducts in alkylated DNA Each method is described by an author who has regularly used the tech­ in his or her own laboratory. Not all the techniques necessarily repre­ nique sent the state-of-the-art-they are, however, all dependable methods that regularly achieve the desired result. Each chapter starts with a brief description of the basic theory behind the technique being described. The main aim of this book, however, is to outline the practical steps necessary for carrying out the method successfully.

Beyond question...the book will be used in many laboratories....[A] fine addition to the...library of publications on immunological methods.-Molecular Immunology

• Medical Research Council Laboratories, Carshalton, UK

  Margaret M. Manson

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