Immunochemical ProtocolsJohn Pound Springer Science & Business Media, 2008 M02 3 - 508 pages This much anticipated second edition provides a user-friendly, up-to-date handbook of reliable immunochemical techniques optimized for molecular biologists. It covers the breadth of relevant established methods from protein blotting and immunoassays through to 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. This book will open the door to a new generation of immunochemical reagents with exciting possibilities. |
Contents
| 1 | |
| 15 | |
Preparation and Testing of Monoclonal Antibodies | 23 |
A Simple HollowFiber Bioreactor for the InHouse Production | 39 |
Screening of Monoclonal Antibodies Using Antigens Labeled | 57 |
Synthesis of Peptides for Use as Immunogens | 69 |
Use of Proteins Blotted to Polyvinylidene Difluoride Membranes | 81 |
IgG Purification | 105 |
Electron MicroscopicSilver Enhancement for Double Labeling | 313 |
Flow Cytometric Analysis of Cell Surface Antigen Density | 319 |
Multiple Immunofluorescence Analysis of Cells Using Flow Cytometry | 337 |
Flow Cytometric Methods of Analyzing Apoptotic Cells | 347 |
Flow Cytometric Detection of ProliferationAssociated Antigens | 355 |
Sorting of Human Peripheral Blood TCell Subsets Using | 365 |
Preparation and Use of Nonradioactive Hybridization Probes | 377 |
Cellular Human and Viral DNA Detection by Nonisotopic In Situ | 385 |
Purification of Monoclonal Antibodies | 113 |
Double Label Immunohistochemistry on Tissue Sections Using | 271 |
Immunogold Probes in Light Microscopy | 283 |
Immunogold Probes in Electron Microscopy | 297 |
Biotinylated Probes in Colony Hybridization | 397 |
Gene Mapping and Cytogenetics | 405 |
Expression of Recombinant Antibody Fusion Proteins in E coli | 419 |
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Common terms and phrases
80 Immunochemical Protocols affinity aliquots alkaline phosphatase analysis anti antibody fragments antigen assay avidin beads binding Biochem biotin biotinylated BrdU buffer cells Centrifuge chain chromatography clones coli column concentration conjugate containing culture described detection diluted Dissolve distilled water ELISA elution enzyme epitope ethanol filter flow cytometry fluorescence fractions fusion gene gold human hybridization hybridoma immunization immunoglobulin Immunol immunolabeling Incubate labeling membrane Methods in Molecular mg/mL microscopy mixture Molecular Biology molecules monoclonal antibodies mouse NaCl nitrocellulose Note overnight PCNA pellet peptide peroxidase phage phagemid plates polyclonal prepared primary antibody probes procedure protein purification PVDF rabbit reaction reagents Resuspend room temperature saline sample SDS-PAGE secondary antibody Section sequences serum sodium sodium azide solution specific staining step Store at 4°C streptavidin substrate supernatant technique tion tissue transfer Tris-HCl tube vector Wash Western blots µg/mL