Tetramers were produced in-house using refolded monomeric, biotinylated pMHC, and streptavidin-PE (Biolegend) at a 1:4 molar ratio. streptavidin-PE was added in 10 steps and incubated for 10 min while shaking at room temperature. Insoluble proteins were removed by brief centrifugation at 13,000 g and 0.05–0.1% sodium azide added for preservation. Tetramers were kept for up to 3 months at 4°C. Cells were stained for CD69 with clones FN50 (Biolegend). Staining for CD45 (clone HI30; Biolegend) was used to distinguish target and effector cells in co-culture assays with U87 cells. Cell viability staining was routinely performed for plate stimulations and U87 co-culture using fixable violet or near-infrared viability dyes (Zombie UV fixable viability kit [Biolegend], Zombie NIR fixable viability kit [Biolegend], eBioscience fixable viability dye eFluor 780 [Invitrogen]). Samples were analysed using a BD X-20 flow cytometer, and data analysis was performed using FlowJo v10 (BD Biosciences).
Clones fn50
Manufactured by BioLegend
The Clones FN50 is a lab equipment product designed for cell culture applications. It functions as a tabletop centrifuge, providing a reliable and consistent method for separating cellular components from liquid samples.
Automatically generated - may contain errors
Lab products found in correlation
X 20 flow cytometer, by BD (2 mentions)
Zombie nir fixable viability kit, by BioLegend (2 mentions)
Streptavidin pe, by BioLegend (2 mentions)
Zombie uv fixable viability kit, by BioLegend (2 mentions)
Ebioscience fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions)
Efluor 780, by Thermo Fisher Scientific (1 mentions)
2 protocols using clones fn50
1
Tetramers for Cell Staining and Analysis
2
Tetramer Staining for Flow Cytometry
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Tetramers were produced in-house using refolded monomeric, biotinylated pMHC and streptavidin-PE (Biolegend) at a 1:4 molar ratio. streptavidin-PE was added in 10 steps and incubated for 10 min while shaking at room temperature. Insoluble proteins were removed by brief centrifugation at 13,000 g and 0.05-0.1 % sodium azide added for preservation. Tetramers were kept for up to 3 months at 4°C. Cells were stained for CD69 with clones FN50 (Biolegend). Staining for CD45 (clone HI30; Biolegend) was used to distinguish target and effector cells in co-culture assays with U87 cells. Cell viability staining was routinely performed for plate stimulations and U87 co-culture using fixable violet or near-infrared viability dyes (Zombie UV fixable viability kit [Biolegend], Zombie NIR fixable viability kit [Biolegend], eBioscience fixable viability dye eFluor 780
[Invitrogen]). Samples were analysed using a BD X-20 flow cytometer and data analysis was performed using FlowJo v10 (BD Biosciences).
[Invitrogen]). Samples were analysed using a BD X-20 flow cytometer and data analysis was performed using FlowJo v10 (BD Biosciences).
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