For analyzing lysosomal acidification, day 3-stimulated naïve CD4+ T cells were treated with 1 μM LysoSensor (L7535, Thermo Fisher Scientific) diluted in prewarmed culture medium and incubated for 30 min at 37°C, followed by washes with ice-cold PBS. LysoSensor fluorescence was quantified by flow cytometry. For examining lysosomal proteolytic activities, cells were treated with DQ-BSA (5 μg/ml; D12050, Thermo Fisher Scientific) diluted in prewarmed culture medium and further incubated at 37°C for 6 hours. After incubation, cells were briefly washed once with ice-cold PBS containing 2% FBS. Fluorescence of cleaved DQ-BSA was analyzed by flow cytometry.
D12050
Manufactured by Thermo Fisher Scientific
The D12050 is a laboratory equipment product from Thermo Fisher Scientific. It is designed for general laboratory use. The core function of the D12050 is to perform basic laboratory tasks, but a detailed description is not available while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using d12050
1
Lysosomal Acidification and Proteolysis in Activated T Cells
2
Quantifying Lysosomal Proteolysis in T Cells
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For studying lysosomal proteolysis, activated naïve CD4+ T cells were treated with DQ-BSA (5 μg/ml; D12050, Thermo Fisher Scientific) diluted in prewarmed medium and incubated at 37°C for 5 hours. Cells were then briefly washed once with ice-cold PBS containing 2% FBS and kept on ice. Fluorescence of cleaved BSA-DQ was analyzed by flow cytometry. For quantitatively analyzing LysoTracker staining, naïve CD4+ T cells were treated with 100 nM LysoTracker (L7528, Thermo Fisher Scientific) diluted in prewarmed medium, incubated for 2 min at 37°C, and then washed with ice-cold PBS containing 2% FBS and kept on ice. LysoTracker fluorescence was analyzed and quantified by flow cytometry.
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