Abstract
A screening method employing immunofluorescence has been developed to judge the amount of specific polypeptides, such as the high lysine protein chymotrypsin inhibitor 2 (CI-2) in the endosperm of barley.
Thin sections or sanded seeds were prepared fixed in 4% paraformaldehyde and labelled with antibody against CI-2 conjugated with fluorescein isothiocyanate. With this technique single seeds can be evaluated without destroying their viability.
The deposition of CI-2 was followed during the grain development period in seeds of Hiproly and of the Hiproly sisterline (CI 4362) both by the immunofluorescence technique and by rocket immunoelectrophoresis. Accumulation of CI-2 started about eight days later in CI 4362 than in Hiproly. The demonstrable amounts of CI-2 in CI 4362 were 20–25% of the amounts found in Hiproly.
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Abbreviations
- BSA:
-
bovine serum albumin
- CI-1:
-
chymotrypsin inhibitor 1
- CI-2:
-
chymotrypsin inhibitor 2
- FITC:
-
fluorescein isothiocyanate
- IgG:
-
immunoglobulin G
- PBS:
-
phosphate buffer saline
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
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Accepted by:H. Klenow, E. Lund andS.O. Andersen
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Rasmussen, U. Immunological screening for specific protein content in barley seeds. Carlsberg Res. Commun. 50, 83 (1985). https://doi.org/10.1007/BF02907139
DOI: https://doi.org/10.1007/BF02907139