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A novel epitope tag for the detection of rabGTPases
Author(s) -
Nagelkerken Bas,
Mohrmann Karin,
Gerez Lisya,
van Raak Marcel,
Leijendekker Richtje,
van der Sluijs Peter
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150181427
Subject(s) - rab , epitope , monoclonal antibody , endosome , epitope mapping , gtpase , linear epitope , antibody , immunocytochemistry , microbiology and biotechnology , biology , conformational epitope , intracellular , cytoplasm , organelle , chemistry , biochemistry , immunology , endocrinology
Rab GTPases are localized on the cytoplasmic surface of most intracellular organelles where they play a role in the regulation of vesicular transport. As it has been difficult to detect endogenous rab proteins by morphological methods, their localizations were often inferred from transfection experiments using epitope‐tagged constructs. Because most of the available epitope tags are only recognitzed by mouse monoclonal antibodies they are often not suitable for double or triple label immunocytochemistry. To overcome this problem, we generated antibodies against a novel 10 amino acid X31 influenza hemagglutin epitope (NH). We here characterized these antibodies and document their utility for detecting early endosomal rab proteins N ‐terminally tagged with the NH decapeptide in morphological and biochemical assays.