Books  Animal & General Biology  Biochemistry & Molecular Biology 

Recombinant Proteins: Detection and Isolation Protocols

Handbook / Manual

Series: Methods in Molecular Biology Volume: 63

By: RS Tuan

470 pages, B/w photos, line illus

Humana Press

Spiralbound | Dec 1996 | #62099 | ISBN: 0896034003
Availability: Usually dispatched within 1 week Details
NHBS Price: £113.00 $146/€124 approx

About this book

Leading researchers and experts present wide-ranging methods for detecting and isolating expressed gene products - recombinant proteins. These state-of-the-art techniques describe a large number of molecular tags and labels, including enzymes, ligand-binding moieties, and immunodetectable molecules. There are also methods to detect interactive proteins and gene expression-mediated alterations in cellular activity, as well as chapters on in situ detection of gene expression. When combined with a companion volume by the same editor, Recombinant Gene Expression Protocols, both volumes guide the research worker through the entire journey of recombinant gene expression.


Part I. Introduction. Overview of Experimental Strategies for the Detection and Isolation of Recombinant Proteins and Their Applications, Rocky S. Tuan. Part II. Detection and Isolation of Expressed Proteins. Reporter Systems, Debyra J. Groskreutz and Elaine T. Schenborn. Detection of Recombinant Protein Based on Reporter Enzyme Activity: Chloramphenicol Acetyltransferase, Peiyu Lee and Dennis E. Hruby. Human Placental Alkaline Phosphatase as a Marker for Gene Expression, Paul Bates and Michael H. Malim. Use of Secreted Alkaline Phosphatase as a Reporter of Gene Expression in Mammalian Cells, Steven R. Kain. Detection of b-Galactosidase and b-Glucuronidase Using Chemiluminescent Reporter Gene Assays, Corinne E. M. Olesen, John J. Fortin, John C. Voyta, and Irena Bronstein. Chemiluminescent Immunoassay for the Detection of Chloramphenicol Acetyltransferase and Human Growth Hormone Reporter Genes, Corinne E. M. Olesen, John C. Voyta, and Irena Bronstein. Detection and Selection of Cultured Cells Secreting Recombinant Product by Soft Agar Cloning and Antibody Overlay, Marylou G. Gibson, Karmen Hodges, and Lauretta Lowther. Detection and Isolation of Recombinant Protein Based on Binding Affinity Reporter: Maltose Binding Protein, Paul Riggs. Detection and Isolation of Recombinant Proteins Based on Binding Affinity of Reporter: Protein A, Stefan Stahl, Per-Ake Nygren, and Mathias Uhlen. Expression and Purification of Recombinant Streptavidin-Containing Chimeric Proteins, Takeshi Sano, Cassandra L. Smith, and Charles R. Cantor. Bacterial Expression, Purification, and Potential Use of His-Tagged GAL4 Fusion Proteins, M. Lienhard Schmitz and Patrick A. Baeuerle. Detection of Expressed Recombinant Protein Based on Multidrug Resistance: P-Glycoprotein, Ursula A. Germann. Detection and Isolation of Recombinant Proteins from Mammalian Cells by Immunoaffinity Chromatography: p53, Jamil Momand and Bahman Sepehrnia. Yeast GAL4 Two-Hybrid System: A Genetic System to Identify Proteins That Interact with a Target Protein, Li Zhu. Alternative Yeast Two-Hybrid Systems: The Interaction Trap and Interaction Mating, Erica A. Golemis and Vladimir Khazak. Detection of Heterologous Gs-Coupled Receptor Activity in LLC-PK1 Cells Based on Expression of Urokinase-Type Plasminogen Activator, Luigi Catanzariti and Brian A. Hemmings. Part III. Detection of Recombinant Gene Expression In Situ. Histochemical and Detection of b-Galactosidase, Ruth Sullivan and Cecilia W. Lo. Combined, Sequential In Situ Hybridization and Immunohistochemistry on the Same Tissue Section, Kenneth J. Shepley and Rocky S. Tuan. Whole Mount In Situ Hybridization to Embryos and Embryonic Tissues, Ronald A. Conlon. Whole-Mount In Situ Hybridization for Developing Chick Embryos Using Digoxygenin-Labeled RNA Probes, Christopher W. Hsu and Rocky S. Tuan. Methods for Double Detection of Gene Expression: Combined In Situ Hybridization and Immunocytochemistry or Histochemistry, Ronald A. Conlon. In Situ PCR: Current Protocol, Omar Bagasra, Muhammad Mukhtar, Farida Shaheen, and Roger J. Pomerantz. Expression and Detection of Green Fluorescent Protein (GFP), Steven R. Kain and Paul Kitts. Part IV. Applications of Recombinant Gene Expression. Transgenic Animals as Bioreactors for Expression of Recombinant Proteins, Jaspal S. Khillan. Artificial Cells for Bioencapsulation of Cells and Genetically Engineered E. coli: For Cell Therapy, Gene Therapy, and Removal of Urea and Ammonia, Thomas M. S. Chang and Satya Prakash. Hyperexpression of a Synthetic Protein-Based Polymer Gene, Henry Daniell, Chittibabu Guda, David T. McPherson, Xiaorong Zhang, Jie Xu, and Dan W. Urry. Transplantation of Cells in an Immunoisolation Device for Gene Therapy, Victoria E. Carr-Brendel, Robin L. Geller, Tracy J. Thomas, Daniel R. Boggs, Susan K. Young, JoAnne Crudele, Laura A. Martinson, David A. Maryanov, Robert C. Johnson, and James H. Brauker. Delivery of Recombinant HIV-1-Directed Antisense and Ribozyme Genes, Georg Sczakiel, Giorgio Palu, and William James. Inducible System Designed for Future Gene Therapy, Yaolin Wang, Bert W. O'Malley, and Sophia Y. Tsai. Human Gene Therapy: Dreams to Realization, Muhammad Mukhtar, Zahida Parveen, and Omar Bagasra. Index.

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