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Ribosome associated proteins of functional ribosomes from murine spermatocytes and round spermatids

ABSTRACT: Murine spermatocytes and round spermatids were isolated by STA-PUT method (J Vis Exp. 2013; (80): 50648.). In order to avoid contamination of nuclear ribosomal subunits and proteins, cytoplasm fractions were extracted by REAP method(BMC Res Notes 3, 294 (2010).) right after cell collection. Immunoprecipitation of functional ribosomes was performed by targeting Rpl36 with the corresponding antibody (Bethyl Laboratories, A305065A). This procedure to capture functional ribosomes was supported by a previous publication (Cell . 2017 Jun 1;169(6):1051-1065.e18.). Rabbit IgG (Cell Signalling Technology, #2729) was used as the negative control. Quantitative mass spectrometry analyses were performed by TMT10plex Isobaric Label Reagents and Kits (Thermo Scientific, 90110).

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Yuh-Shan Jou

PROVIDER: MSV000091932 | MassIVE | Sat May 13 01:54:00 BST 2023

SECONDARY ACCESSION(S): PXD042202

REPOSITORIES: MassIVE

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