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Study of lipid rafts by gel filtration combined with preliminary staining with fluorescently labeled antibodies

  • M. P. Krutikova
  • G. I. Krotov
  • V. G. Zgoda
  • A. V. Filatov
Article

Abstract

A novel procedure for detecting raft-associated proteins by gel filtration was developed. Cells were stained with fluorescently labeled protein-specific antibodies, lyzed in a nonionic detergent and gel-filtered on Sepharose 4B. Proteins were identified with the help of fluorescently labeled antibodies. Staining of cells with fluorescently labeled antibodies prior to cell lysis significantly simplified the identification procedure. The potentialities of the new approach are demonstrated in the example of several surface proteins associated with lipid rafts either constitutively or in the course of cell activation.

Keywords

Free Volume Lipid Raft Cholera Toxin Supplement Series Brij 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

DRM

detergent-resistant membranes

FITC

fluorescein isothiocyanate

PAGE

polyacrylamide gel

PBS

phosphate-buffered saline

CT

cholera toxin B-subunit

GPI

glycosylphosphatidylinositol

PMSF

phenylmethylsulfonyl fluoride

mAb

monoclonal antibody

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Copyright information

© Pleiades Publishing, Ltd. 2007

Authors and Affiliations

  • M. P. Krutikova
    • 1
  • G. I. Krotov
    • 1
  • V. G. Zgoda
    • 2
  • A. V. Filatov
    • 1
  1. 1.National Research Center “Institute of Immunology, Federal Medico-Biological Agency of Russia”MoscowRussia
  2. 2.Orekhovich Institute of Biomedical ChemistryRussian Academy of Medical SciencesMoscowRussia

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