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Heat solubilization of Grl1p. Isolated granules (30 μg protein/aliquot) in 1% TX-100 were incubated at 95°C for 30 min, with or without previous exposure to 1 mM calcium. Samples were then centrifuged at 17,000 × g for 30 min. Fifty percent of the supernatant fractions and 100% of the pellets were analyzed by SDS-PAGE (17.5% polyacrylamide) and visualized by Coomassie blue.
