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Fig. 2. Subcellular fractionation of RBL cell lysate. Fractions from a continuous sucrose gradient were collected from the top and assayed for (A) Syt III (anti Syt III serum 1:1000 dilution), Syt II (monoclonal antibodies 1:1000 dilution) and G{alpha}i2 (affinity-purified AS10 antibodies, 1 µg/ml) immunoreactivities. (B) ß-hexosaminidase activity (presented as OD read at 405 nm) ({blacksquare}) and histamine content ({square}). (C) Protein (•) and sucrose density ({circ}). (D) The light density fractions of the gradient were probed for Syt III (polyclonal anti Syt III serum 1: 1000 dilution), Annexin II (monoclonal 1: 5000 dilution), EEA1 (polyclonal anti EEA1 serum 1: 1000 dilution) and syntaxin 7 (polyclonal anti syntaxin 7 antibodies 1: 500 dilution) immunoreactivities. Data represent one of three separate experiments.





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