Membrane:Article Title: Coptis chinensis extracellular vesicles loaded with CA1-siRNA promote endothelial repair and stent restenosis therapy by regulating the PADI2 and NF-κB pathway.
Article Snippet: .. After sealing, the membrane was incubated overnight with primary antibodies at 4 °C: anti-CA1 (13198-2-AP, Proteintech, China), anti-IL-6 (16806-1-AP, Proteintech, China), anti-IL-1β (K002095P, Solarbio, China), antiTNF-α (K002095P, Solarbio, China), anti-alpha SMA (BM0002, Boster, China), anti-MMP9 (BM4089, Boster, China), anti-IκBα (abs131168, absinthe, China), antiIκBα phosphorylation (abs172314, absinthe, China), anti-p65 (BF8005, Affinity, China), anti-p-p65 (bs-0271R, BIOSS, China), and anti-β-actin (66009-1-Ig, Proteintech, China). .. After washing, incubate the membrane with horseradish peroxidase conjugated secondary antibody for 1 h. The signals were detected using a chemiluminescence imaging system (Millipore, USA) and LumiQ universal ECL substrate (SB-WB012, Solarbio, Shanghai), and quantified using ImageJ software, normalized to β-actin levels.
Article Title: Coptis chinensis extracellular vesicles loaded with CA1-siRNA promote endothelial repair and stent restenosis therapy by regulating the PADI2 and NF-κB pathway
Article Snippet: .. After sealing, the membrane was incubated overnight with primary antibodies at 4 °C: anti-CA1 (13198-2-AP, Proteintech, China), anti-IL-6 (16806-1-AP, Proteintech, China), anti-IL-1β (K002095P, Solarbio, China), anti-TNF-α (K002095P, Solarbio, China), anti-alpha SMA (BM0002, Boster, China), anti-MMP9 (BM4089, Boster, China), anti-IκBα (abs131168, absinthe, China), anti- IκBα phosphorylation (abs172314, absinthe, China), anti-p65 (BF8005, Affinity, China), anti-p-p65 (bs-0271R, BIOSS, China), and anti-β-actin (66009-1-Ig, Proteintech, China). .. After washing, incubate the membrane with horseradish peroxidase conjugated secondary antibody for 1 h. The signals were detected using a chemiluminescence imaging system (Millipore, USA) and LumiQ universal ECL substrate (SB-WB012, Solarbio, Shanghai), and quantified using ImageJ software, normalized to β-actin levels.
Incubation:Article Title: Coptis chinensis extracellular vesicles loaded with CA1-siRNA promote endothelial repair and stent restenosis therapy by regulating the PADI2 and NF-κB pathway.
Article Snippet: .. After sealing, the membrane was incubated overnight with primary antibodies at 4 °C: anti-CA1 (13198-2-AP, Proteintech, China), anti-IL-6 (16806-1-AP, Proteintech, China), anti-IL-1β (K002095P, Solarbio, China), antiTNF-α (K002095P, Solarbio, China), anti-alpha SMA (BM0002, Boster, China), anti-MMP9 (BM4089, Boster, China), anti-IκBα (abs131168, absinthe, China), antiIκBα phosphorylation (abs172314, absinthe, China), anti-p65 (BF8005, Affinity, China), anti-p-p65 (bs-0271R, BIOSS, China), and anti-β-actin (66009-1-Ig, Proteintech, China). .. After washing, incubate the membrane with horseradish peroxidase conjugated secondary antibody for 1 h. The signals were detected using a chemiluminescence imaging system (Millipore, USA) and LumiQ universal ECL substrate (SB-WB012, Solarbio, Shanghai), and quantified using ImageJ software, normalized to β-actin levels.
Article Title: Coptis chinensis extracellular vesicles loaded with CA1-siRNA promote endothelial repair and stent restenosis therapy by regulating the PADI2 and NF-κB pathway
Article Snippet: .. After sealing, the membrane was incubated overnight with primary antibodies at 4 °C: anti-CA1 (13198-2-AP, Proteintech, China), anti-IL-6 (16806-1-AP, Proteintech, China), anti-IL-1β (K002095P, Solarbio, China), anti-TNF-α (K002095P, Solarbio, China), anti-alpha SMA (BM0002, Boster, China), anti-MMP9 (BM4089, Boster, China), anti-IκBα (abs131168, absinthe, China), anti- IκBα phosphorylation (abs172314, absinthe, China), anti-p65 (BF8005, Affinity, China), anti-p-p65 (bs-0271R, BIOSS, China), and anti-β-actin (66009-1-Ig, Proteintech, China). .. After washing, incubate the membrane with horseradish peroxidase conjugated secondary antibody for 1 h. The signals were detected using a chemiluminescence imaging system (Millipore, USA) and LumiQ universal ECL substrate (SB-WB012, Solarbio, Shanghai), and quantified using ImageJ software, normalized to β-actin levels.
Phospho-proteomics:Article Title: Coptis chinensis extracellular vesicles loaded with CA1-siRNA promote endothelial repair and stent restenosis therapy by regulating the PADI2 and NF-κB pathway.
Article Snippet: .. After sealing, the membrane was incubated overnight with primary antibodies at 4 °C: anti-CA1 (13198-2-AP, Proteintech, China), anti-IL-6 (16806-1-AP, Proteintech, China), anti-IL-1β (K002095P, Solarbio, China), antiTNF-α (K002095P, Solarbio, China), anti-alpha SMA (BM0002, Boster, China), anti-MMP9 (BM4089, Boster, China), anti-IκBα (abs131168, absinthe, China), antiIκBα phosphorylation (abs172314, absinthe, China), anti-p65 (BF8005, Affinity, China), anti-p-p65 (bs-0271R, BIOSS, China), and anti-β-actin (66009-1-Ig, Proteintech, China). .. After washing, incubate the membrane with horseradish peroxidase conjugated secondary antibody for 1 h. The signals were detected using a chemiluminescence imaging system (Millipore, USA) and LumiQ universal ECL substrate (SB-WB012, Solarbio, Shanghai), and quantified using ImageJ software, normalized to β-actin levels.
Article Title: Coptis chinensis extracellular vesicles loaded with CA1-siRNA promote endothelial repair and stent restenosis therapy by regulating the PADI2 and NF-κB pathway
Article Snippet: .. After sealing, the membrane was incubated overnight with primary antibodies at 4 °C: anti-CA1 (13198-2-AP, Proteintech, China), anti-IL-6 (16806-1-AP, Proteintech, China), anti-IL-1β (K002095P, Solarbio, China), anti-TNF-α (K002095P, Solarbio, China), anti-alpha SMA (BM0002, Boster, China), anti-MMP9 (BM4089, Boster, China), anti-IκBα (abs131168, absinthe, China), anti- IκBα phosphorylation (abs172314, absinthe, China), anti-p65 (BF8005, Affinity, China), anti-p-p65 (bs-0271R, BIOSS, China), and anti-β-actin (66009-1-Ig, Proteintech, China). .. After washing, incubate the membrane with horseradish peroxidase conjugated secondary antibody for 1 h. The signals were detected using a chemiluminescence imaging system (Millipore, USA) and LumiQ universal ECL substrate (SB-WB012, Solarbio, Shanghai), and quantified using ImageJ software, normalized to β-actin levels.
other:
Article Title: M1-Type Macrophages Secrete TNF-α to Stimulate Vascular Calcification by Upregulating CA1 and CA2 Expression in VSMCs
Article Snippet: The proteins were transferred to a PVDF membrane (Millipore, Burlington, MA, USA), which was then blocked with 5% nonfat milk for 2 h. The membrane was incubated overnight at 4°C with primary antibodies against CA1, CA2 (13198-2-AP/16961-1-AP, Proteintech) or GAPDH (AF0502, Elabscience).
|