To capture ubiquitin chains from cell extracts, 10 μl of packed beads were incubated with 1 mg cell extract protein. For enrichment of ubiquitylated proteins from phagosomal proteins after magnetic beads isolation, 60 μg phagosomal extract was incubated with 10 μl of packed beads. Samples were incubated for 16 h at 4°C at end‐over‐end mixing and beads were washed three times with IP washing buffer 50 mM Tris, 150 mM NaCl, 1% Triton‐X 100. If no other treatment was required samples were denatured by LDS sample buffer. Beads were transferred to SpinX tubes (Corning) to collect sample for SDS/PAGE.
Spinx tubes
Manufactured by Corning
SpinX tubes are a type of laboratory equipment designed for centrifugation. They are cylindrical containers used to hold samples during the centrifugation process. The core function of SpinX tubes is to facilitate the separation of different components within a liquid sample through the application of centrifugal force.
Automatically generated - may contain errors
3 protocols using spinx tubes
1
Ubiquitin Chain Enrichment from Cell Extracts
2
Tumor Protein Extraction and Cytokine Analysis
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Tumors were excised, weighed, and ground with a pestle in 1:20 (wt/vol) of tumor in Tissue Protein Extraction Reagent (Thermo Fisher) supplemented with Halt Protease Inhibitor Mixture (Thermo Fisher) and 5 mM ethylenediaminetetraacetic acid (EDTA). Samples were incubated for 30 min at 4 °C, debris was pelleted by centrifugation at 10,000 × g, and supernatants were filtered through Corning Costar Spin-X tubes. The LEGENDplex mouse anti-virus response panel (13-plex) (BioLegend) was used following vendor instructions and analyzed with BD FACS LSR Fortessa.
3
Ubiquitin Chain Enrichment from Cell Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
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To capture ubiquitin chains from cell extracts 10 µl of packed beads were incubated with 1mg cell extract protein. For enrichment of ubiquitylated proteins from phagosomal proteins after magnetic beads isolation, 60 µg phagosomal extract was incubated with 10 µl of packed beads. Samples were incubated for 16 h at 4°C at end-over-end mixing and beads were washed 3 times with IP washing buffer 50 mM Tris, 150 mM NaCl, 1% Triton-X 100. If no other treatment was required slurry was denatured by LDS sample buffer. Beads were transferred to SpinX tubes (Corning) to collect sample for SDS/PAGE.
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