Eμ-TCL1 splenocytes, human CLL PBMCs (hCLL), and sorted T-cell subsets from mice and humans were cultured at 1x106/mL for 24 hours in 96 well plates. Concentrations of inhibitors and stimulants are listed in figure legends. Anti-IgM (Jackson ImmunoResearch, West Grove, PA), anti-CD3, anti-CD28 (BioLegend) and MTM (Enzo Life Sciences, Farmingdale, NY) were commercially sourced, while MTMox32E was synthesized in house(26 (link)). Secreted cytokines were measured with ELISA MAX Standard Set Mouse or Human kits according to the manufacturer’s protocol (BioLegend). Cellular viability was determined by resuspending cells in cRPMI with 0.1mM resazurin (Sigma Aldrich), incubating for four hours at 37°C, and measuring fluorescence (ex:560nm, em:590nm) on a SpectraMax M5 plate reader (Molecular Devices, Sunnyvale, CA)(27 (link)).
Elisa max standard set mouse or human kits
Manufactured by BioLegend
The ELISA MAX Standard Set Mouse or Human kits are lab equipment designed for the quantitative measurement of the levels of specific proteins in biological samples. These kits provide the necessary components, including pre-coated plates, detection antibodies, and standards, to perform enzyme-linked immunosorbent assay (ELISA) procedures.
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2 protocols using elisa max standard set mouse or human kits
1
Cytokine Secretion and Viability Assays
2
Cytokine Secretion and Viability Assays
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Eμ-TCL1 splenocytes, human CLL PBMCs (hCLL), and sorted T-cell subsets from mice and humans were cultured at 1x106/mL for 24 hours in 96 well plates. Concentrations of inhibitors and stimulants are listed in figure legends. Anti-IgM (Jackson ImmunoResearch, West Grove, PA), anti-CD3, anti-CD28 (BioLegend) and MTM (Enzo Life Sciences, Farmingdale, NY) were commercially sourced, while MTMox32E was synthesized in house(26 (link)). Secreted cytokines were measured with ELISA MAX Standard Set Mouse or Human kits according to the manufacturer’s protocol (BioLegend). Cellular viability was determined by resuspending cells in cRPMI with 0.1mM resazurin (Sigma Aldrich), incubating for four hours at 37°C, and measuring fluorescence (ex:560nm, em:590nm) on a SpectraMax M5 plate reader (Molecular Devices, Sunnyvale, CA)(27 (link)).
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