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Identification of sarkosyl-insoluble proteins that accumulate in the brains of aged mice.

ABSTRACT: Five young (2-month-old) and five aged (24-month-old) mouse brains were bisected along the midline to separate the right and left hemispheres, then snap-frozen in liquid nitrogen. The frozen tissues were pulverized using a Multi-Beads Shocker (Yasui Kikai) and stored at -150°C until further use. The brain powder was homogenized in sarkosyl buffer (1% sarkosyl, 10 µg/mL RNase, 1 mM PMSF, 1 mM EDTA in PBS) using a Potter-Elvehjem homogenizer at 800 rpm. After a 10-minute incubation on ice, the homogenate was centrifuged at 15,000 g for 5 minutes at 4°C to separate the supernatant and pellet (P1). The supernatant was centrifuged again at 15,000 g for 5 minutes at 4°C, followed by a third centrifugation at 15,000 g for 15 minutes at 4°C to obtain the final supernatant (S1) and a pellet. S1 was then subjected to ultracentrifugation at 100,000 g for 1 hour at 4°C, resulting in a supernatant (S2) and a pellet. This pellet was resuspended in sarkosyl buffer using a vortex mixer and centrifuged again at 100,000 g for 1 hour at 4°C. The supernatant was discarded, and the final pellet (P2), representing the sarkosyl-insoluble fraction, was retained.

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Shigeo Murata

PROVIDER: PXD060043 | JPOST Repository | Mon Aug 04 00:00:00 BST 2025

REPOSITORIES: jPOST

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			Action	DRS
	210218Kominato100,000g_ppt.mzid	Mzid
	210218KominatoA4.mzML	Mzml
	210218KominatoA4.raw	Raw
	210218KominatoB4.mzML	Mzml
	210218KominatoB4.raw	Raw

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Identification of sarkosyl-insoluble proteins that accumulate in the brains of aged mice.

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