Milk:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0.
Lysis:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0.
Protease Inhibitor:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0.
Activation Assay:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0.
Cell Isolation:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0.
Software:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0.
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