Total proteins were extracted from cells and quantified with the BCA method, which were separated using the 12% SDS-PAGE, followed by transferred to the PVDF membrane. Following blocking, primary antibodies against RET (1:1000, AF6120, Affinity, USA), p-AKT (1:1000, AF0016, Affinity, USA), AKT (1:1000, AF6259, Affinity, USA), mTOR (1:1000, AF6308, Affinity, USA), p-mTOR (1:1000, AF3308, Affinity, USA), and GAPDH (1:10000, 1094-1-AP, Proteintech, USA) were added to be cultured for 12 h at 4 ℃, followed by incubation with the secondary antibody (1:6000, 7074, CST, USA) and incubated for 60 min. Lastly, the ECL solution was added for exposure and the expression level was quantified with the Image J software.
Af6259
Manufactured by Affinity Biosciences
Sourced in United States
AF6259 is a specialized laboratory instrument designed for quantitative analysis. It utilizes a proven detection method to provide precise and reliable measurements. The core function of this product is to support research and testing activities in a wide range of scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using af6259
1
Quantitative Western Blot Analysis
2
Protein Expression Analysis via Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
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EPCs were collected to extract total proteins, followed by quantified using the BCA method. After conducting the separation with the 12% SDS-PAGE, proteins were transferred to the PVDF membrane, which was then blocked in the 5% skim milk. The primary antibodies against p-PI3K (1:1000, AF3241, Affinity, USA), PI3K (1:1000, AF6241, Affinity, USA), p-AKT (1:1000, AF0016, Affinity, USA), AKT (1:1000, AF6259, Affinity, USA), eNOS (1:1000, AF6247, Affinity, USA), p-eNOS (1:1000, AF3247, Affinity, USA), and GAPDH (1:10000, 10494-1-AP, Proteintech, USA) were added, followed by incubation with the secondary antibody (1:6000, 7074, CST, USA). After incubation for 1 h, the ECL solution was added for exposure and the expression level was quantified with the Image J software.
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