Summary
Immunogold labelling has been used to study the cellular and subcellular localization of myrosinase (β-thioglucosidase, EC 3.2.3.1), using LR-White acrylic resin and ultrahin sections from four different species of Brassicaceae;Brassica napus L.,Sinapis alba L.,Raphanus sativus L., and B.oleracea L. For immunolabelling, a polyclonal antibody raised in rabbit against a highly purified myrosinase fromSinapis alba was used. Western blots showed that the antiserum was specific against myrosinase from these species. Ultrathin sections were sequentially incubated with anti-myrosinase antiserum and with secondary antibodies conjugated with colloidal gold. Gold label was present in typical myrosin cells both in radicles and in cotyledons when observed in the electron microscope. The intracellular localization showed that myrosinase was present in myrosin grains in the myrosin cells in all four species of Brassicaceae.
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Abbreviations
- BSA:
-
bovine serum albumin
- PBS:
-
phosphate buffered saline
- PBS-T:
-
PBS with 0.5% v/v Tween-20
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Thangstad, O.P., Evjen, K. & Bones, A. Immunogold-EM localization of myrosinase in Brassicaceae. Protoplasma 161, 85–93 (1991). https://doi.org/10.1007/BF01322721
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DOI: https://doi.org/10.1007/BF01322721