Western Blotting for Protein Analysis

Western blotting was performed using a gel documentation system (iBrightCL1000, Invitrogen and Image Lab Software version 3.0), and a standard protocol as previously described (Zhang et al., 2019 (link)). The primary antibodies were anti-cyclin dependent kinase inhibitor 2A (P16) (80772, Cell Signaling Technology, United States), anti-cyclin dependent kinase inhibitor 1A (P21) (2947, Cell Signaling Technology), P53 (21083, Signalway Antibody, United States), PTEN (ab31392, abcam, United Kingdom), anti-phosphorylated-AKT (p-AKT) (4060, Cell Signaling Technology) and anti-AKT (4691, Cell Signaling Technology); Cleaved caspase-3 (29034, Signalway Antibody), Bcl-2 (ab196495, abcam), vascular endothelial growth factor (VEGF, ab52917, abcam), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (5174, Cell Signaling Technology); TSG101 (14497, Proteintech, United States), CD63 (25682, Proteintech), CD81 (66866, Proteintech) and horseradish peroxidase-conjugated were secondary antibodies (Biosharp, China).

Free full text: Click here

Sun L., Zhu W., Zhao P., Zhang J., Lu Y., Zhu Y., Zhao W., Liu Y., Chen Q, & Zhang F. (2020). Down-Regulated Exosomal MicroRNA-221 – 3p Derived From Senescent Mesenchymal Stem Cells Impairs Heart Repair. Frontiers in Cell and Developmental Biology, 8, 263.

Publication 2020

iBrightCL1000 ab31392 anti-AKT Cleaved caspase-3 anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) TSG101 horseradish peroxidase-conjugated

Antibodies Antibody Bcl 2 Caspase 3 Glyceraldehyde 3 phosphate dehydrogenase Horseradish peroxidase Pten Tsg101 Vegf

Corresponding Organization : Nanjing Medical University

Other organizations : Changzhou No.2 People's Hospital, Zhongda Hospital Southeast University

Top 5 similar protocols

Variable analysis

independent variables

None specified

dependent variables

Expression levels of: P16, P21, P53, PTEN, p-AKT, AKT, Cleaved caspase-3, Bcl-2, VEGF

control variables

Western blotting protocol and gel documentation system (iBrightCL1000, Invitrogen and Image Lab Software version 3.0)
GAPDH, TSG101, CD63, CD81 as loading controls

controls

Positive controls: None specified
Negative controls: None specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!