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

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Biochemistry (Moscow) Supplement Series A: Membrane and Cell Biology Aims and scope

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.

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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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Authors and Affiliations

  1. National Research Center “Institute of Immunology, Federal Medico-Biological Agency of Russia”, Kashirskoe sh. 24/2, Moscow, 115478, Russia

    M. P. Krutikova, G. I. Krotov & A. V. Filatov

  2. Orekhovich Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Pogodinskaya ul. 10, Moscow, 119121, Russia

    V. G. Zgoda

Authors
  1. M. P. Krutikova
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  2. G. I. Krotov
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  3. V. G. Zgoda
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  4. A. V. Filatov
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Correspondence to M. P. Krutikova.

Additional information

Original Russian Text © M.P. Krutikova, G.I. Krotov, V.G. Zgoda, A.V. Filatov, 2007, published in Biologicheskie Membrany, 2007, Vol. 24, No. 4, pp. 299–308.

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Krutikova, M.P., Krotov, G.I., Zgoda, V.G. et al. Study of lipid rafts by gel filtration combined with preliminary staining with fluorescently labeled antibodies. Biochem. Moscow Suppl. Ser. A 1, 219–227 (2007). https://doi.org/10.1134/S199074780703004X

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  • DOI: https://doi.org/10.1134/S199074780703004X

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