Three-dimensional immunofluorescent double labelling using polyclonal antibodies derived from the same species: enterocytic colocalization of chylomicrons with Golgi apparatus.
Histochem Cell Biol
; 129(6): 779-84, 2008 Jun.
Article
em En
| MEDLINE
| ID: mdl-18299879
ABSTRACT
Double immunolabelling is a useful technique to determine cellular colocalization of proteins, but is prone to false-positive staining because of cross-reactivity between antibodies. In this study, we established a simple and quick method to demonstrate the immunofluorescent double labelling with two rabbit-derived polyclonal antibodies. The principle used was to establish a dilution of primary antibody for the first protein of interest, which would only be detectable following biotin-avidin amplification. Thereafter, the second protein of interest was assessed via standard secondary antibody detection, ensuring no cross-reactivity with the first protein antibody-antigen complex. We successfully demonstrated the three-dimensional colocalization of enterocytic apolipoprotein B, an equivocal marker of intestinal lipoproteins with Golgi apparatus. Colocalization of apo B and Golgi apparatus (75.2 +/- 8.5%) is consistent with the purported mode of secretion of these macromolecules.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Quilomícrons
/
Enterócitos
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Complexo de Golgi
/
Anticorpos
Idioma:
En
Ano de publicação:
2008
Tipo de documento:
Article