Production of Polyclonal Antisera -- Raising Polyclonal Antibodies Using Nitrocellulose-Bound Antigen -- Production and Characterization of Antibodies Against Synthetic Peptides -- Preparation and Testing of Monoclonal Antibodies to Recombinant Proteins -- A Simple Hollow-Fiber Bioreactor for the "In-House" Production of Monoclonal Antibodies -- Screening of Monoclonal Antibodies Using Antigens Labeled with Acetylcholinesterase -- Synthesis of Peptides for Use as Immunogens -- Use of Proteins Blotted to Polyvinylidene Difluoride Membranes as Immunogens -- Purification of Glycoproteins and Their Use as Immunogens -- IgG Purification -- Purification of Monoclonal Antibodies -- Production of Bispecif ic and Trispecific F(ab)2 and F(ab)3 Antibody Derivatives -- Preparation of Cytotoxic Antibody-Toxin Conjugates -- Immunoaffinity Purification and Quantification of Antibody-Toxin Conjugates -- Competitive ELISA -- Epitope Mapping -- Coupling of Antibodies with Biotin -- Preparation of Avidin Conjugates -- Enhanced Chemiluminescence Immunoassay -- Nonradioactive Methods for Visualization of Protein Blots -- Colloidal Gold Staining and Immunoprobing on the Same Western Blot -- Erasure of Western Blots After Autoradiographic or Chemiluminescent Detection -- Colloidal Gold Staining and Immunodetection in 2D Protein Mapping -- Enzyme-Antienzyme Method for Immunohistochemistry -- Immunohistochemical Detection of Bromodeoxyuridine-Labeled Nuclei for In Vivo Cell Kinetic Studies -- Cell Kinetic Studies Using a Monoclonal Antibody to Bromodeoxyuridine -- Immunohistochemical Detection of Cells in the Division Cycle Using Antibodies to Proliferating Cell Nuclear Antigen (PCNA) -- Double Label Immunohistochemistry on Tissue Sections Using Alkaline Phosphatase and Peroxidase Conjugates -- Preparation of Colloidal Gold Probes -- Immunogold Probes in Light Microscopy -- Immunogold Probes in Electron Microscopy -- Electron Microscopic-Silver Enhancement for Double Labeling with Antibodies Raised in the Same Species -- Flow Cytometric Analysis of Cell Surface Antigen Density -- Multiple Immunofluorescence Analysis of Cells Using Flow Cytometry -- Flow Cytometric Methods of Analyzing Apoptotic Cells -- Flow Cytometric Detection of Proliferation-Associated Antigens, PCNA and Ki-67 -- Sorting of Human Peripheral Blood T-Cell Subsets Using Immunomagnetic Beads -- Preparation and Use of Nonradioactive Hybridization Probes -- Cellular Human and Viral DNA Detection by Nonisotopic In Situ Hybridization -- Biotinylated Probes in Colony Hybridization -- Nonisotopic In Situ Hybridization -- Expression of Recombinant Antibody Fusion Proteins in E. coli -- Expression of Antibody Fusion Proteins in Mammalian Cells -- Antibody Screening of Bacteriophage ? gt-11 DNA Expression Libraries -- Display of Antibody Chains on Filamentous Bacteriophage -- Production of Human Fab Antibody Fragments from Phage Display Libraries -- Antibodies from Phage Display Libraries as Immunochemical Reagents.

This much anticipated second edition of the best selling Immunological Protocols provides a user-friendly, up-to-date handbook of reliable immunochemical techniques optimized for ready use by molecular biologists. It covers the breadth of relevant established methods-from protein blotting and immunoassays through the visualization of cellular antigens and in situ hybridization-each with their latest refinements. Protocols for the production and purification of important classes of immunochemical reagents are also provided, including "conventional" and recombinant antibodies, fusion proteins, and their various conjugates. The second edition of Immunochemical Protocols brings many previously specialized techniques within the grasp of all laboratory workers. It remains, like its predecessor, "an excellent laboratory manual and reference" and opens the door to the productive use of a new generation of immunochemical reagents with exciting possibilities.