Protein Expression Analysis by Western Blot
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Corresponding Organization :
Other organizations : Xuzhou Central Hospital
Variable analysis
- MiR-142-5p mimic
- MiR-142-5p inhibitor
- Corresponding negative control
- Protein levels of FOXO1
- Protein levels of FOXO3
- Protein levels of Bcl-2 interacting mediator of cell death (Bim)
- Protein levels of procaspase 3
- Protein levels of active caspase 3
- Protein concentration determined using the BCA Assay Kit
- 10%-12% sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE)
- Blotting onto polyvinylidene fluoride (PVDF) membranes
- Blocking in 5% defatted milk in Tris-buffered saline with Tween (TBST) for 1 h
- Incubation with primary antibodies overnight at 4°C
- Incubation with horseradish peroxidase (HRP)-conjugated antibody for 2 h
- GAPDH used as a loading control
- Protein bands visualized with enhanced chemiluminescence (ECL)
- Not explicitly mentioned
- Corresponding negative control
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