Membrane:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
Protein Extraction:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
Saline:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
Sterility:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
Zeta Potential Analyzer:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
Western Blot:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
Enzyme-linked Immunosorbent Assay:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
Flow Cytometry:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
Protease Inhibitor:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
Detection Assay:Article Title: Drug-free Immunotherapeutic Biomimetic Nanoparticles for Treating Triple-Negative Breast Cancer.
Article Snippet: .. The following materials and reagents were used in this study: Membrane Protein Extraction Kit (ProteoExtract), acetone, chloroform, methanol, DMSO, Tween 20, phosphate-buffered saline (PBS), RPMI-1640, DMEM, Eukitt(R) Quick-hardening mounting medium, 25 mm Sterile Syringe Filters, 0.02 μm PVDF, and cholesterol from Sigma-Aldrich-Merck; 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (PE-Rhod) from Avanti Polar Lipids, Inc.; dialysis tubes (12−14 kDa and 1000 kDa) from Repligen; Dynamic Light Scattering (DLS), ZetaSizer Nano, and disposable cuvettes for zeta potential measurements from Malvern Instruments; rabbit antiCD11b (MAB11242−100) for Western blotting, and DuoSet ELISA kits for TNF-α, TGF-β, IFN-γ and IL-6 from R&D Systems; rabbit antiCD68 (ab125212), rabbit anti-CD163 (ab182422), goat antirabbit IgG-HRP (ab6721), hematoxylin and eosin kit, and DAB substrate kit from Abcam; flow cytometry antibodies: anti-CD45-FITC (BLG157608), anti-CD11b-Alexa Fluor 647 (BLG-101218), anti-F4/80APC-Cy7 (BLG-157315), anti-Ly6G-BV711 (BLG-127643), antiLy6C-PE (BLG-128008), anti-CD3-Spark Blue 574 (BLG-100276), anti-CD8a-Alexa Fluor 594 (BLG-100758), anti-CD4-BV510 (BLG100449), anti-FoxP3-Pacific Blue (BLG-126410), anti-CD19-PE-Cy7 (BLG-152418), and Zombie Violet Viability Kit from BioLegend; semi microvolume disposable polystcytokinesyrene cuvettes for size measurements and MycoStrip Mycoplasma Detection Kit from Tamar Ltd.; 5X Sample Buffer from A2S; Trans-Blot Turbo Mini PVDF membrane, TC20 Automated Cell Counter and slides, and Clarity Western ECL Substrate from Bio-Rad Laboratories; Quick Coating Solution from AngioProteomie; NanoAssemblr Benchtop and Microfluidic Cartridge from Precision Nanosystems (Cytiva); Infinite M Plex multimode microplate reader from Tecan; Pierce BCA kit,1Step TMB ELISA Substrate Solutions, Vybrant DiD Cell-Labeling Solution, Halt protease inhibitor cocktail, Quant-iT Endotoxin Detection Assay Kit (Q32892) and wheat germ agglutinin-Alexa Fluor 488 from ThermoFisher Scientific; μ-slide 0.4 was purchased from ibidi, and Endothelial Cell Medium with supplements from ScienCell; InVivomAb antimouse PD-1 (CD279) (BE0146−5A) from BioXCell. .. Murine J7741.A macrophages (ATCC) were cultured in high-glucose DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
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