Techniques in HIV research

著者

書誌事項

Techniques in HIV research

edited by Anna Aldovini and Bruce D. Walker

Stockton Press, c1990

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  • :us
  • :uk
  • :uk

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注記

Includes bibliographical references and index

内容説明・目次

巻冊次

:uk ISBN 9780333518069

内容説明

A manual that details the techniques of electrofusion and electroporation by researchers who were the first to show that the platelet membrane glycoproteins GP IIb and GP IIIa are associated in a complex. The interest in electrofusion was triggered by the potential of this technique to produce monoclonal antibodies with desired specificity in a more efficient way. Realizing the need for more information and together with Carl Borrebaeck, Inger Hagen arranged the First International Workshop on "Electrofusion in Hybriddoma Technology", Lysebu 1988. This book aims to show how to produce monoclonal antibodies with improved electrofusion techniques, detailing protocols for electrofusion experiments with different cell fusion partners, describing the new electrofusion chamber allowing electrofusion with very few cells, and discussing the potential of directed electrofusion.

目次

  • Electroinjection and electrofusion in hypo-osmolar solution
  • electroporation of human lymphoid cells - methods and application
  • formation of hybridomas secreting human monoclonal antibodies with mouse - human fusion partners
  • generating immortalized immunoglobulin - secreting human lymphocytes by recominat DNA tehnology
  • B-cell hybridoma production by avidin-biotin mediated electrofusion.
巻冊次

:uk ISBN 9780333535400

内容説明

A comprehensive compilation of research techniques necessary for investigating the virology, immunology and molecular biology of HIV-1. Protocols are also provided which represent state of the art approaches to a wide spectrum of HIV related issues.

目次

  • Part 1 Antibody detection in patient's serum: ELISA assay
  • western blot analysis. Part 2 Virus detection in pathological specimen: p24 assay
  • polymerase chain reaction (PCR)
  • in situ hybridization
  • southern blot analysis. Part 3 Virus isolation and production: isolation from periperal blood
  • isolation from tissues
  • virus production in stable cell lines. Part 4 Virus titration: on MT4 cells
  • on HeLa cells
  • on C81-66
  • on SUP T1. Part 5 Virus detection in cell lines: reverse transcriptase assay
  • p24 assay
  • immunofluorescence (IF)
  • cythopathic effect (CPE)
  • radioimmunoprecipitation (RIP). Part 6 Virus purification and storage. Part 7 Proviral DNA cloning: recombinant DNA libraries
  • polymerase chain reaction (PCR). Part 8 Tranfection of biologically active molecular clones: DEAE dextran
  • electroporation
  • CaPO4 transfection
  • lipofection
  • selection of stably transfected cell lines. Part 9 Host immune response: neutralizing antibodies
  • ADCC/CMC
  • cytotoxity - CTL
  • proliferative response. Part 10 Cytokine assays. Part 11 Antiviral drugs assays. Appendices: source of reagents
  • biosafey issues.

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