Mass Spectrometry Analysis of C. elegans ISW-1::GFP
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ABSTRACT: The germ cell nuclear extracts of ISW-1::GFP were incubated with GFP Nanoab Agarose Beads (GNA-25-500, NuoyiBio) for 6 hours at 4°C. The mixture was then washed five times with lysis buffer (20 mM Tris-HCl pH 7.6, 300 mM NaCl, 1.5 mM MgCl2, 20 mM KCl, 1% NP-40, 1 mM DTT, and protease inhibitors). Immunoprecipitated proteins were eluted by boiling in 1% SDS and 25 mM Tris buffer (pH 7.4) for 5 minutes. The eluted proteins were precipitated with 20% trichloroacetic acid (TCA) at 4°C for 1 hour, followed by centrifugation at 14,000 rpm for 10 minutes at 4°C. The precipitate was then washed once with 10% TCA and twice with cold acetone (14,000 rpm for 10 minutes at 4°C). The final protein pellet was collected and digested with trypsin for mass spectrometry analysis.
ORGANISM(S): Caenorhabditis Elegans
SUBMITTER:
Jinmin Gao
PROVIDER: PXD067287 | iProX | Wed Aug 13 00:00:00 BST 2025
REPOSITORIES: iProX
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