Aqueous Normal-phase Chromatography:Article Title: ABCC9 knockdown attenuates isoproterenol-induced myocardial hypertrophy by inhibiting the PI3K/AKT signaling pathway
Article Snippet: An anti-ABCC9 antibody (cat. no. 40229) was purchased from Signalway Antibody LLC. .. Antibodies against B-cell lymphoma 2 protein (Bcl-2; cat. no. 15071T), Bcl-2-associated X protein (Bax; cat. no. 2772T), caspase-3 (cat. no. 9662S), cleaved caspase-3 (cat. no. 9661S), phosphorylated (p)-PI3K (cat. no. 17366T), PI3K (cat. no. 4292S), p-AKT (cat. no. 9271T), AKT (cat. no. 4691T) and β-actin (cat. no. 3700S) were purchased from Cell Signaling Technology, Inc. Atrial natriuretic peptide (ANP; cat. no. ab189921) and brain natriuretic peptide (BNP; cat. no. ab236101) were purchased from Abcam. .. GAPDH (cat. no. sc-32233) and secondary antibodies were purchased from Santa Cruz Biotechnology, Inc., including mouse anti-rabbit IgG-HRP (cat. no. sc-2357) and mouse IgGκ light chain binding protein conjugated to HRP (cat. no. sc-516102).
Immunohistochemical staining:
Staining:
Blocking Assay:Article Title: Oral Administration of Astrocyte-Targeted Natural Antioxidants Suppress NOX4-Driven Neuroinflammation and Restore Hippocampal Neurogenesis in MPTP-Induced Parkinson’s Disease Mouse Model
Article Snippet: Protein concentrations were measured, and equal amounts were subjected to SDS-PAGE and transferred onto PVDF membranes (Cytiva, Marlborough, MA, USA). .. After blocking with TBST containing 5% BSA, membranes were incubated overnight (4 °C) with primary antibodies: 4-Hydroxynonenal (4-HNE; 1:1000, Bioss, Beijing, China), Bcl-2-Associated X Protein (BAX; 1:1000, Cell signaling Technology), B-Cell Lymphoma 2 (Bcl-2; 1:1000, Cell signaling Technology), cleaved caspase-3 (CC3; 1:500, Cell Signaling Technology), DCX (1:1000, Abcam), GFAP (1:1000, Merck Millipore, Burlington, MA, USA), Ionized calcium binding adaptor molecule 1 (Iba-1; 1:200, Wako, Osaka, Japan), MPO (1:500, R&D systems), OPN (1:100, R&D Systems), NeuN (1:500, Abcam), NOX4 (1:100, Novus, Centennial, CO, USA), PSD95 (1:1000, Invitrogen, Waltham, MA, USA), Synaptophysin (1:20,000, Novus), TIM23 (1:1000, Proteintech, Chicago, IL, USA), Tom20 (1:100, Santa Cruz Biotechnology), Total OXPHOS (1:250, Abcam) and Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH; 1:10,000, Cell Signaling). .. After washing, membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (Vector Laboratories, Cell Signaling) for 2 h, and immunoreactive bands were visualized using enhanced chemiluminescence reagents (BS ECL Plus Kit; Biosesang, Yongin-si, Gyeonggi-do, Republic of Korea).
Incubation:Article Title: Oral Administration of Astrocyte-Targeted Natural Antioxidants Suppress NOX4-Driven Neuroinflammation and Restore Hippocampal Neurogenesis in MPTP-Induced Parkinson’s Disease Mouse Model
Article Snippet: Protein concentrations were measured, and equal amounts were subjected to SDS-PAGE and transferred onto PVDF membranes (Cytiva, Marlborough, MA, USA). .. After blocking with TBST containing 5% BSA, membranes were incubated overnight (4 °C) with primary antibodies: 4-Hydroxynonenal (4-HNE; 1:1000, Bioss, Beijing, China), Bcl-2-Associated X Protein (BAX; 1:1000, Cell signaling Technology), B-Cell Lymphoma 2 (Bcl-2; 1:1000, Cell signaling Technology), cleaved caspase-3 (CC3; 1:500, Cell Signaling Technology), DCX (1:1000, Abcam), GFAP (1:1000, Merck Millipore, Burlington, MA, USA), Ionized calcium binding adaptor molecule 1 (Iba-1; 1:200, Wako, Osaka, Japan), MPO (1:500, R&D systems), OPN (1:100, R&D Systems), NeuN (1:500, Abcam), NOX4 (1:100, Novus, Centennial, CO, USA), PSD95 (1:1000, Invitrogen, Waltham, MA, USA), Synaptophysin (1:20,000, Novus), TIM23 (1:1000, Proteintech, Chicago, IL, USA), Tom20 (1:100, Santa Cruz Biotechnology), Total OXPHOS (1:250, Abcam) and Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH; 1:10,000, Cell Signaling). .. After washing, membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (Vector Laboratories, Cell Signaling) for 2 h, and immunoreactive bands were visualized using enhanced chemiluminescence reagents (BS ECL Plus Kit; Biosesang, Yongin-si, Gyeonggi-do, Republic of Korea).
Binding Assay:Article Title: Oral Administration of Astrocyte-Targeted Natural Antioxidants Suppress NOX4-Driven Neuroinflammation and Restore Hippocampal Neurogenesis in MPTP-Induced Parkinson’s Disease Mouse Model
Article Snippet: Protein concentrations were measured, and equal amounts were subjected to SDS-PAGE and transferred onto PVDF membranes (Cytiva, Marlborough, MA, USA). .. After blocking with TBST containing 5% BSA, membranes were incubated overnight (4 °C) with primary antibodies: 4-Hydroxynonenal (4-HNE; 1:1000, Bioss, Beijing, China), Bcl-2-Associated X Protein (BAX; 1:1000, Cell signaling Technology), B-Cell Lymphoma 2 (Bcl-2; 1:1000, Cell signaling Technology), cleaved caspase-3 (CC3; 1:500, Cell Signaling Technology), DCX (1:1000, Abcam), GFAP (1:1000, Merck Millipore, Burlington, MA, USA), Ionized calcium binding adaptor molecule 1 (Iba-1; 1:200, Wako, Osaka, Japan), MPO (1:500, R&D systems), OPN (1:100, R&D Systems), NeuN (1:500, Abcam), NOX4 (1:100, Novus, Centennial, CO, USA), PSD95 (1:1000, Invitrogen, Waltham, MA, USA), Synaptophysin (1:20,000, Novus), TIM23 (1:1000, Proteintech, Chicago, IL, USA), Tom20 (1:100, Santa Cruz Biotechnology), Total OXPHOS (1:250, Abcam) and Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH; 1:10,000, Cell Signaling). .. After washing, membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (Vector Laboratories, Cell Signaling) for 2 h, and immunoreactive bands were visualized using enhanced chemiluminescence reagents (BS ECL Plus Kit; Biosesang, Yongin-si, Gyeonggi-do, Republic of Korea).
|