The hippocampi were homogenized in lysis buffer (RIPA) on ice for 30 min and subsequently centrifuged at 12000 rpm for 5 min at 4 °C. Protein concentrations were determined using a bicinchoninic acid protein assay kit (Thermo Fisher Scientific, USA). Equivalent amounts of proteins were processed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot as previously described [29 (link)]. The primary antibodies used in this experiment were Kir4.1 (1:1000, Abcam), AKT (1:1000, CST), Arc (1:1000, CST), PSD95 (1:1000, CST), GluN2A (1:1000, Abcam), GluA1 (1:1000, Abcam), and β-actin (1:1000, CST). Band intensity was densitometrically quantified using Image J software.
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Hippocampal Protein Expression Analysis
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Actin
Antibodies
Assay
Bicinchoninic acid
Buffer
Glun2a
Hippocampi
Proteins
Ripa
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
Western blot
Corresponding Organization :
Other organizations : Anhui University of Traditional Chinese Medicine, Zhejiang Chinese Medical University
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Variable analysis
independent variables
- Homogenization of hippocampi in lysis buffer (RIPA) on ice for 30 min
- Protein concentrations
- Band intensity of Kir4.1, AKT, Arc, PSD95, GluN2A, GluA1, and β-actin proteins
- Centrifugation at 12000 rpm for 5 min at 4 °C
- Use of bicinchoninic acid protein assay kit (Thermo Fisher Scientific, USA)
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot as previously described [29]
- No positive or negative controls were explicitly mentioned in the protocol.
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