Staining:Article Title: A TROP2/Claudin Program Mediates Immune Exclusion to Impede Checkpoint Blockade in Breast Cancer
Article Snippet: .. Cells were stained with Viability Ghost Dye (Tonbo Biosciences, 13-0870) at 4 °C for 10 mins, followed by anti-CD16/32 blocking (Tonbo Bioscience, 70-0161-U500) at 4 °C for 10 min. For surface proteins, cells were stained with anti-CD45 (BioLegend, 103108), anti-CD3 (BioLegend, 100209), anti-CD8a (BioLegend, 100714), anti-TIM3 (BioLegend, 119703), anti-CD44 (BioLegend, 103041), anti-CD62L (BioLegend, 104410), and anti-PD1 (BD Biosciences, 563059) in FACS buffer (2% FBS in PBS). .. For cytokine staining, cells were treated with anti-CD3/CD28 (Thermo Fisher, 11452D) at 37 °C overnight and then with BD GolgiPlug (BD Biosciences, 550583) at 37 °C for 5 h. Following surface staining, cells were permeabilized using BD Cytofix/Cytoperm kit (BD Biosciences, 554714), and stained with anti-Granzyme B (Invitrogen, 48-8898-82).
Article Title: TROP2/claudin program mediates immune exclusion to impede checkpoint blockade in breast cancer.
Article Snippet: .. Cells were stained with Viability Ghost Dye (Tonbo Biosciences, 13–0870) at 4°C for 10 min, followed by antiCD16/32 blocking (Tonbo Bioscience, 70–0161 U500) at 4°C for 10 min. For surface proteins, cells were stained with anti- CD45 (BioLegend, 103108), anti- CD3 (BioLegend, 100209), anti- CD8a (BioLegend, 100714), anti- TIM3 (BioLegend, 119703), anti- CD44 (BioLegend, 103041), anti- CD62L (BioLegend, 104410), and antiPD- 1 (BD Biosciences, 563059) in FACS (fluorescenceactivated cell sorting) buffer (2% FBS (fetal bovine serum) in PBS). .. For cytokine staining, cells were treated with anti- CD3/CD28 (Thermo Fisher, 11 452D) at 37°C overnight and then with BD GolgiPlug (BD Biosciences, 550583) at 37°C for 5 hours.
Blocking Assay:Article Title: A TROP2/Claudin Program Mediates Immune Exclusion to Impede Checkpoint Blockade in Breast Cancer
Article Snippet: .. Cells were stained with Viability Ghost Dye (Tonbo Biosciences, 13-0870) at 4 °C for 10 mins, followed by anti-CD16/32 blocking (Tonbo Bioscience, 70-0161-U500) at 4 °C for 10 min. For surface proteins, cells were stained with anti-CD45 (BioLegend, 103108), anti-CD3 (BioLegend, 100209), anti-CD8a (BioLegend, 100714), anti-TIM3 (BioLegend, 119703), anti-CD44 (BioLegend, 103041), anti-CD62L (BioLegend, 104410), and anti-PD1 (BD Biosciences, 563059) in FACS buffer (2% FBS in PBS). .. For cytokine staining, cells were treated with anti-CD3/CD28 (Thermo Fisher, 11452D) at 37 °C overnight and then with BD GolgiPlug (BD Biosciences, 550583) at 37 °C for 5 h. Following surface staining, cells were permeabilized using BD Cytofix/Cytoperm kit (BD Biosciences, 554714), and stained with anti-Granzyme B (Invitrogen, 48-8898-82).
Article Title: TROP2/claudin program mediates immune exclusion to impede checkpoint blockade in breast cancer.
Article Snippet: .. Cells were stained with Viability Ghost Dye (Tonbo Biosciences, 13–0870) at 4°C for 10 min, followed by antiCD16/32 blocking (Tonbo Bioscience, 70–0161 U500) at 4°C for 10 min. For surface proteins, cells were stained with anti- CD45 (BioLegend, 103108), anti- CD3 (BioLegend, 100209), anti- CD8a (BioLegend, 100714), anti- TIM3 (BioLegend, 119703), anti- CD44 (BioLegend, 103041), anti- CD62L (BioLegend, 104410), and antiPD- 1 (BD Biosciences, 563059) in FACS (fluorescenceactivated cell sorting) buffer (2% FBS (fetal bovine serum) in PBS). .. For cytokine staining, cells were treated with anti- CD3/CD28 (Thermo Fisher, 11 452D) at 37°C overnight and then with BD GolgiPlug (BD Biosciences, 550583) at 37°C for 5 hours.
FACS:Article Title: A TROP2/Claudin Program Mediates Immune Exclusion to Impede Checkpoint Blockade in Breast Cancer
Article Snippet: .. Cells were stained with Viability Ghost Dye (Tonbo Biosciences, 13-0870) at 4 °C for 10 mins, followed by anti-CD16/32 blocking (Tonbo Bioscience, 70-0161-U500) at 4 °C for 10 min. For surface proteins, cells were stained with anti-CD45 (BioLegend, 103108), anti-CD3 (BioLegend, 100209), anti-CD8a (BioLegend, 100714), anti-TIM3 (BioLegend, 119703), anti-CD44 (BioLegend, 103041), anti-CD62L (BioLegend, 104410), and anti-PD1 (BD Biosciences, 563059) in FACS buffer (2% FBS in PBS). .. For cytokine staining, cells were treated with anti-CD3/CD28 (Thermo Fisher, 11452D) at 37 °C overnight and then with BD GolgiPlug (BD Biosciences, 550583) at 37 °C for 5 h. Following surface staining, cells were permeabilized using BD Cytofix/Cytoperm kit (BD Biosciences, 554714), and stained with anti-Granzyme B (Invitrogen, 48-8898-82).
Article Title: TROP2/claudin program mediates immune exclusion to impede checkpoint blockade in breast cancer.
Article Snippet: .. Cells were stained with Viability Ghost Dye (Tonbo Biosciences, 13–0870) at 4°C for 10 min, followed by antiCD16/32 blocking (Tonbo Bioscience, 70–0161 U500) at 4°C for 10 min. For surface proteins, cells were stained with anti- CD45 (BioLegend, 103108), anti- CD3 (BioLegend, 100209), anti- CD8a (BioLegend, 100714), anti- TIM3 (BioLegend, 119703), anti- CD44 (BioLegend, 103041), anti- CD62L (BioLegend, 104410), and antiPD- 1 (BD Biosciences, 563059) in FACS (fluorescenceactivated cell sorting) buffer (2% FBS (fetal bovine serum) in PBS). .. For cytokine staining, cells were treated with anti- CD3/CD28 (Thermo Fisher, 11 452D) at 37°C overnight and then with BD GolgiPlug (BD Biosciences, 550583) at 37°C for 5 hours.
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