Cytotoxicity was determined using ARP1-GFPLuc as target tumor cells. Briefly, ARP1-GFPLuc cells were cultured with CARHigh T and CARLow T cells at different ratios in RPMI 1640 culture medium supplemented with 3% human serum (Sigma-Aldrich) and 1% penicillin/streptomycin in Nunc 96-well round bottom plates (Thermo Fisher Scientific). After 24 hours, luminescence was measured using the Bright-Glo Luciferase Assay System (Promega) according to the manufacturer’s instructions. IFN-γ, TNFα, and IL-2 cytokine production was quantified using BD Immunoassay enzyme-linked immunosorbent assay (ELISA) reagents (BD Biosciences) following the manufacturer’s protocol.
Rpmi 1640 culture
Manufactured by Merck Group
RPMI 1640 is a cell culture medium commonly used for the in vitro cultivation of a variety of cell types, including human and animal cells. It provides a balanced salt solution and essential nutrients required for cell growth and proliferation. The medium is formulated to maintain a stable pH and support the metabolic needs of cultured cells.
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Lab products found in correlation
2 protocols using rpmi 1640 culture
1
Cytotoxicity Assay of CAR T Cells
2
Mass Spectrometry Sample Preparation
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All materials, unless otherwise stated, were purchased from Sigma-Aldrich (St. Louis, MO).
RPMI1640 culture medium, fetal bovine serum, glutamine (GlutaMAX I), sodium pyruvate, Optimem (Opti-MEM I), MEM nonessential amino acids, and cell culture CD-CHO medium were purchased, as Gibco Life Technologies products, from Thermo Fisher Scientific (Waltham, MA).
α-Cyano-4-hydroxycinnamic acid matrix (CHCA), Peptide Calibration Standard II, 3 × MTP target insert polished plain 1.0, and an MTP sheet metal adaptor 1.0 mm were purchased from Bruker Daltonics (Bremen, Germany).
RPMI1640 culture medium, fetal bovine serum, glutamine (GlutaMAX I), sodium pyruvate, Optimem (Opti-MEM I), MEM nonessential amino acids, and cell culture CD-CHO medium were purchased, as Gibco Life Technologies products, from Thermo Fisher Scientific (Waltham, MA).
α-Cyano-4-hydroxycinnamic acid matrix (CHCA), Peptide Calibration Standard II, 3 × MTP target insert polished plain 1.0, and an MTP sheet metal adaptor 1.0 mm were purchased from Bruker Daltonics (Bremen, Germany).
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