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After vigorous shaking, the fungal material was pelleted, the supernatant discarded and the mycelium dried in a concentrator (room temperature, 60 min; Eppendorf, Hamburg, Germany).
Insoluble material was pelleted by centrifugation for 30 minutes at 21.000 g.
Cells were freeze-thawed three times and insoluble material was pelleted.
Detergent-insoluble material was pelleted by microcentrifugation (13,000 rpminutesnutes, 4°C).
Insoluble material was pelleted and the resulting supernatant transferred to a new microfuge tube and evaporated to dryness under low heat in a rotary evaporator.
Cell debris and insoluble material was pelleted by centrifugation at 4°C for 20,000 g for 20 min. The supernatant was stored at −20°C.
Similar(17)
Membrane materials were pelleted by ultracentrifugation at 4°C (2 times 70 min at 285000 g).
Insoluble materials were pelleted by centrifugation at 15000× g for 20 min at 4°C. 10 30 µg of protein extracts were used for direct Western blotting.
The debris and insoluble materials were pelleted down at 15,000 rev/min using a Beckman JA20 rotor.
The gel slice along with the 3 mm paper was incubated in 100 μL deionized water for 30 min, boiled for 15 min in a tightly capped eppendorf tube, and the gel materials were pelleted by centrifugation.
containing 1% Triton X-100, 150 mM NaCl, 1 mM NaF, 0.1 mM phenylmethylsulfonyl fluoride, 5 μg/ml pepstatin A and 10 μg/ml each of leupeptin and aprotinin (lysis buffer) and insoluble materials were pelleted by centrifugation at 12 000 × g for 30 min at 4 °C.
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