PD-L1 siRNA (cat# L-015836–01-0005) and control siRNA were purchased from GE Dharmacon and transfection experiments were performed using lipofectamine 3000 (Invitrogen) per manufacturer’s protocol and described previously.39 Briefly, cells were seeded in 6-well plate at 1 × 105 per well seeding density. lipofectamine 3000 reagent and siRNA were diluted separately in Opti-MEM media and mixed together. Lipid-siRNA complex was further incubated for 5 min and then added to respective wells. Post 24 and 48 h of transfection, cells were harvested by trypsinization and western blotting was performed to assess PD-L1 knockdown.
Pd l1 sirna
Manufactured by Horizon Discovery
Sourced in Germany
PD-L1 siRNA is a laboratory tool designed for gene silencing. It targets the expression of the PD-L1 (Programmed Death-Ligand 1) gene, which is involved in the regulation of the immune response. This siRNA can be used in cell-based assays and research applications to study the role of PD-L1 in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Control sirna, by Horizon Discovery (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Human recombinant interferon beta, by R&D Systems (1 mentions)
Bms 345541, by Merck Group (1 mentions)
Clone 913 429, by R&D Systems (1 mentions)
Mouse anti human β actin, by Cell Signaling Technology (1 mentions)
Goat anti mouse igg secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Methylthiazolyldiphenyl tetrazolium bromide mtt, by Merck Group (1 mentions)
Calcein am, by Thermo Fisher Scientific (1 mentions)
Soluble recombinant human trail, by Enzo Life Sciences (1 mentions)
2 protocols using pd l1 sirna
1
PD-L1 Knockdown via siRNA Transfection
2
Chemotherapeutic Agents and Immunomodulators
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Chemotherapeutics used for this study were cisplatin (Teva; Ulm, Germany), 5-fluorouracil (Medac; Hamburg, Germany) and gemcitabine (Hexal; Holzkirchen, Germany). Methylthiazolyldiphenyl-tetrazolium bromide (MTT) was obtained from Sigma Aldrich (St. Louis; MO, USA), human recombinant interferon beta from R&D System (New York; NY, USA), the PD-1 inhibitor nivolumab from Bristol-Myers (Anagni, Italy). Calcein-AM was purchased from Thermo Fisher (Eugene, USA), soluble, recombinant human TRAIL from Enzo Life Science (Paris, France). Primary mouse monoclonal antibody against PD-1, clone 913429 was obtained from R&D System (Wiesbaden, Germany), anti-human B7-H1/PD-L1 monoclonal antibody, clone 130021 from R&D System (Minneapolis, USA). Knock-down experiments were performed using PD-L1-siRNA (Dharmacon; Freiburg, Germany, Cat. Nr. L-015836-01-0010), RELA-siRNA for NF-κB (Dharmacon; Cat. Nr. L-003533-00-0005) and scrambled RNA (Dharmacon; Cat. Nr. D-001810-20-0005). Antibodies used for immunoblotting were mouse anti-human β-actin (Cell Signaling; Danvers, MA, USA), mouse anti-human-PD-1 (R&D System), mouse anti-human-PD-L1 (Clone # 130021; R&D System) and mouse anti-human-RELA/NF-κB p65 (Clone #532301; R&D System). The goat anti-mouse IgG secondary antibody was purchased from Santa Cruz Biotechnology (Heidelberg, Germany). NF-κB inhibitor—BMS-345541 was obtained from Sigma Aldrich.
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