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M9640689.TXT
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1996-03-04
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Document 0689
DOCN M9640689
TI Detection of intracellular HIV-1 Rev protein by flow cytometry.
DT 9604
AU Rigg RJ; Dando JS; Escaich S; Plavec I; Bohnlein E; Progenesys, Palo
Alto, CA 94304, USA.
SO J Immunol Methods. 1995 Dec 27;188(2):187-95. Unique Identifier :
AIDSLINE MED/96144820
AB The Rev trans-activator protein plays a pivotal role in human
immunodeficiency virus type 1 (HIV-1) replication by allowing expression
of the viral structural proteins. We have developed a protocol to
quantitatively assay intracellular steady state levels of Rev Ag (Rev
wild type and RevM10 proteins) by flow cytometry. Three fixation and
permeabilization techniques were compared. These protocols varied in the
magnitude of the signal which could be detected, and in the ability to
distinguish between Rev Ag positive and negative populations. This
technology is applicable to a variety transduced or transfected cell
types (species, lineage), and for cell lines and primary cells acutely
infected with HIV-1. The assay is therefore a valuable tool both to
analyze Rev protein expression levels in HIV-infected cells and to
optimize delivery of the dominant-negative RevM10 gene for clinical gene
therapy applications. In addition, a second, independent intracellular
protein (HIV-Tat) has been detected using the same approach.
DE Animal Antibodies, Monoclonal Cell Line Cells, Cultured Comparative
Study Flow Cytometry/*METHODS Gene Products, rev/*ANALYSIS Gene
Products, tat/ANALYSIS Hela Cells/VIROLOGY Human
HIV-1/*CHEMISTRY/PHYSIOLOGY Lymphocytes/VIROLOGY Mice Permeability
Reproducibility of Results Tissue Fixation Transfection 3T3
Cells/VIROLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).