NDLI: Three-dimensional immunofluorescent double labelling using polyclonal antibodies derived from the same species: enterocytic colocalization of chylomicrons with Golgi apparatus

Content Provider	Springer Nature Link
Author	Takechi, R. Galloway, S. Pallebage Gamarallage, M. M. S. Johnsen, R. D. Mamo, J. C. L.
Copyright Year	2008
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.
Starting Page	779
Ending Page	784
Page Count	6
File Format	PDF
ISSN	09486143
Journal	Histochemistry and Cell Biology
Volume Number	129
Issue Number	6
e-ISSN	1432119X
Language	English
Publisher	Springer-Verlag
Publisher Date	2008-02-26
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Double immunofluorolabelling Three-dimensional colocalization Polyclonal antibodies Intestinal Golgi apparatus Chylomicron Medicine/Public Health Anatomy
Content Type	Text
Resource Type	Article
Subject	Cell Biology Histology Molecular Biology Medical Laboratory Technology

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