Protein Extraction and Quantification from HepG2 and Huh7 Cells
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Corresponding Organization :
Other organizations : Zhejiang Chinese Medical University, University of Chinese Academy of Sciences, Zhejiang Cancer Hospital
Variable analysis
- Cell line (HepG2 and Huh7)
- Total protein amount
- Levels of β-actin and hnRNPA2B1 proteins
- RIPA Lysis Buffer (Beyotime Institute of Biotechnology) for cell lysis
- Centrifugation at 1,200 × g at 4°C for 10 min
- BCA assay for protein quantification
- SDS-PAGE electrophoresis at 60 V
- Transfer to polyvinylidene difluoride membrane
- Blocking with 5% (w/v) non-fat dry milk at 37°C for 1 h
- Primary antibodies against β-actin (1:2,000 dilution) and hnRNPA2B1 (1:1,000 dilution)
- HRP-conjugated secondary antibodies (1:5,000 dilution)
- Chemiluminescence detection using Pierce (Thermo Fisher Scientific, Inc.) and Tanon 5200 Imaging system
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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