R-phycoerythrin as a fluorescent label for immunolocalization of bound atrazine residues

• Published in: The journal of histochemistry and cytochemistry Vol. 39; no. 7; pp. 921 - 926
• Main Authors: Sohn, G, Sautter, C
• Format: Journal Article
• Language: English
• Published: Los Angeles, CA Histochemical Soc 01.07.1991; SAGE Publications; Histochemical Society
• Subjects: Atrazine - analysis; Biological and medical sciences; Cell structures and functions; Fluorescent Antibody Technique; Fluorescent Dyes; Fundamental and applied biological sciences. Psychology; Molecular and cellular biology; Phycoerythrin; Plants - analysis; Immunofluorescence; Localization; Fluorescent labelling; Phycoerythrin
• ISSN: 0022-1554; 1551-5044
• DOI: 10.1177/39.7.1865109

We established a highly sensitive immunofluorescence procedure for localizing bound atrazine in the aquatic macrophytes Elodea canadensis and E. densa. The technique included biotin-labeled anti-rabbit IgG as a first enhancement step and R-phycoerythrin (R-PE) coupled to streptavidin for fluorescent labeling as a second improvement on the procedure. A comparison with the conventional indirect immunofluorescence method confirmed the superior results of the R-PE approach. The use of atrazine-free plants (grown in charcoal-filtered water) and a variety of other controls excluded both contaminating atrazine and nonspecific incubation constituents as sources of tissue staining. Pre-incubations to block nonspecific binding sites proved to be unnecessary in this system. The highly sensitive procedure described here might be a useful tool for the localization of tissue-bound pesticides in general and possibly of other haptens as well.