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Immunofluorescent quantitation of chloroplast proteins
Author(s) -
Leech Rachel M.,
Marrison Joanne L.
Publication year - 1996
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1996.10061169.x
Subject(s) - chloroplast , rubisco , immunofluorescence , in situ , biology , fluorescence , microbiology and biotechnology , chemistry , biochemistry , photosynthesis , optics , antibody , gene , organic chemistry , physics , immunology
Summary Using scanning light microscopy software to detect and measure immunofluorescence in leaf sections Rubisco concentration in situ in chloroplasts has been accurately determined throughout development. The fluorescence measurements were calibrated by comparison with values for Rubisco accumulation obtained from rocket immuno‐ electrophoresis profiles of soluble protein from isolated cells and from chloroplasts using a purified sample of Rubisco as the standard. It has been shown that in situ immunofluorescence can be used for cytoquantitation of proteins within individual chloroplasts to a sensitivity of 1fg and also for the comparison of the protein levels in adjacent chloroplasts and cells. Several important applica‐ tions of this new technique are discussed.