Western Blot Analysis of AKT Proteins

Total protein was extracted from cells using radioimmunoprecipitation assay buffer (Beyotime, China) and the protein was separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, as previously described [33 (link)]. The antibodies used were as follows: anti-AKT3 (Abcam, Cambridge, MA, USA; ab2157); anti-p-AKT (Abcam; ab8805); anti-P-AKT (Abcam; ab8805; ab192623); and anti-GAPDH (Abcam; ab9485). Original Western Blot images were provided in Supplementary file 1.

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Zhou Y., Cai W, & Lu H. (2022). Overexpression of microRNA-145 enhanced docetaxel sensitivity in breast cancer cells via inactivation of protein kinase B gamma-mediated phosphoinositide 3-kinase -protein kinase B pathway. Bioengineered, 13(4), 11310-11320.

Publication 2022

radioimmunoprecipitation assay buffer ab2157 ab8805 ab192623 ab9485

Akt3 Antibodies Buffer Gapdh Protein Radioimmunoprecipitation assay Sodium dodecyl sulfate polyacrylamide gel electrophoresis Western blot

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Variable analysis

independent variables

Radioimmunoprecipitation assay buffer used to extract total protein from cells

dependent variables

Protein expression levels of AKT3
Phosphorylation levels of AKT (p-AKT)

control variables

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis used to separate the proteins
GAPDH used as a loading control

controls

Positive control: Original Western Blot images provided in Supplementary file 1

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