To assess virus replication, mice were euthanized and blood, tumors, and normal tissues were collected and snap-frozen. Mouse plasma was prepared by high-speed centrifugation of EDTA anticoagulated whole blood (13,000 rpm for 10 min at 4°C in a table-top microcentrifuge). Frozen tumors and normal tissues were weighed and suspended in ice-cold homogenization buffer (PBS supplemented with 1× antibiotic-antimycotic [Thermo Fisher Scientific] and 1× Halt protease-phosphatase inhibitor [Thermo Fisher Scientific]) before being transferred to Matrix A tubes (MP Bio, Santa Ana, CA) and homogenized using the FastPrep-24 lysis system (MP Bio) for 20 s at 4 min per second. Tissue homogenates were then subjected to two freeze-thaw cycles, divided into aliquots, and stored at −80°C. Two additional aliquots that were not subjected to freeze thaws were prepared for virus recovery or cytokine analysis by MSD-ECL.
Matrix a tubes
Manufactured by MP Biomedicals
Matrix A tubes are laboratory consumables designed for various applications. They are made of high-quality materials and are suitable for a range of scientific purposes.
Automatically generated - may contain errors
2 protocols using matrix a tubes
1
Characterizing Viral Replication in Mice
2
Bacterial Lipid Extraction and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Bacterial pellets were suspended in 300μL HBSS and frozen at -80°C until lipids extraction. These samples were added with 5 μL of internal standard mixture (Deuterium-labeled compounds, 400 ng/mL) and crushed with 2x30 sec shaking at 5 m/s (Matrix A tubes, FastPrep, MP Biomedical). 300μL of cold methanol was added, the tubes were centrifuged at 1016 × g for 15 min (4°C) and the supernatants were stored at -80°C until extraction. Lipids were extracted on solid phase HLB 96 wells plates (OASIS HLB, Waters) conditioned with 500μL methanol and 500μL 90:10 water/methanol. Samples were brought to 2mL with water and loaded at one drop per 2 s, then washed with 500μL 90:10 water/methanol and dried under aspiration. Lipids were eluted with 750μL methanol and evaporated twice under N2, then suspended in 10 μL methanol. The quantification of C14-asparagine was performed by the MetaToul Lipidomics facility (Inserm UMR1048, Toulouse, France), using an in-house quantification assay by high-performance liquid chromatography/tandem mass spectrometry analysis [36 (link)].
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