UID:
almahu_9949251212102882
Format:
1 online resource (XIII, 480 p.)
Edition:
1st ed. 1992.
ISBN:
1-59259-497-2
Series Statement:
Methods in Molecular Biology, 10
Content:
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.
Note:
Bibliographic Level Mode of Issuance: Monograph
,
Production of Polyclonal Antisera -- Efficient Elution of Purified Proteins from Polyvinylidene Difluoride Membranes (Immobilon) After Transfer from SDS-PAGE and Their Use as Immunogens -- Raising Polyclonal Antibodies Using Nitrocellulose-Bound Antigen -- Synthesis of Peptides for Use as Immunogens -- Production and Characterization of Antibodies Against Synthetic Peptides -- Preparation and Testing of Monoclonal Antibodies to Recombinant Proteins -- Screening of Monoclonal Antibodies Using Antigens Labeled with Acetylcholinesterase -- Purification of Immunoglobulin G (IgG) -- Epitope Mapping -- Enzyme-Antienzyme Method for Immunohistochemistry -- Double Label Immunohistochemistry on Tissue Sections Using Alkaline Phosphatase and Peroxidase Conjugates -- Immunohistochemical Detection of Bromodeoxyuridine-Labeled Nuclei for In Vivo Cell Kinetic Studies -- Avidin–Biotin Technology -- Avidin–Biotin Technology -- Immunochemical Applications of Avidin-Biotin Technology -- Preparation of Gold Probes -- Immunogold Probes for Light Microscopy -- Immunogold Probes in Electron Microscopy -- Electron Microscopic Silver Enhancement for Double Labeling with Antibodies Raised in the Same Species -- Quantitative and Qualitative Immunoelectrophoresis -- Rocket Immunoelectrophoresis Technique or Electroimmunodiffusion -- Crossed Immunoelectrophoresis -- Crossed Immunoaffinoelectrophoresis -- Immunodetection of Proteins by Western Blotting -- Erasable Western Blots -- Colloidal Gold Staining and Immunoprobing on the Same Western Blot -- Colloidal Gold Staining and Immunodetection in 2-D Protein Mapping -- Fluorescent Protein Staining on Nitrocellulose with Subsequent Immunodetection of Antigen -- Competitive ELISA -- Twin-Site ELISAs for fos and myc Oncoproteins Using the AMPAK System -- Preparation of Cytotoxic Antibody-Toxin Conjugates -- Immunoaffinity Purification and Quantification of Antibody-Toxin Conjugates -- An Immuno-Slot-Blot Assay for Detection and Quantitation of Alkyldeoxyguanosines in DNA -- Production of Monoclonal Antibodies for the Detection of Chemically Modified DNA -- Sensitive Competitive Enzyme-Linked Immunoassay for Quantitation of Modified Bases in DNA -- An Immunochemical Assay for Detecting Transition of B-DNA to Z-DNA -- Cell Sorting Using Immunomagnetic Beads -- Cell Preparation for Flow Cytometry -- Preparation of Rat Lung Cells for Flow Cytometry -- The Isolation of Rat Hepatocytes for Flow Cytometry -- Flow Cytometric Analysis of Cells Using an Antibody to a Surface Antigen -- Multiple Immunofluorescence Analysis of Cells Using Flow Cytometry -- Cell Kinetic Studies Using a Monoclonal Antibody to Bromodeoxyuridine -- Production and Use of Nonradioactive Hybridization Probes -- Cellular Human and Viral DNA Detection by Nonisotopic In Situ Hybridization -- Chromosomal Mapping of Genes by Nonisotopic In Situ Hybridization -- Nonisotopic In Situ Hybridization -- Biotinylated Probes in Colony Hybridization -- Screening of ?gt11 cDNA Libraries Using Monoclonal Antibodies -- Expression of Foreign Genes in Mammalian Cells Using an Antibody Fusion System.
,
English
Additional Edition:
ISBN 0-89603-204-3
Language:
English
