Western Blot Analysis of Mechanosensors

Western blotting was performed as previously described [20 (link)]. The blots were incubated overnight with primary antibodies targeting β-actin (Santa Cruz), Piezo1 (Poteintech), NFAT1 (Santa Cruz), NFAT2 (Santa Cruz), NFAT3 (Santa Cruz), NFAT4 (Santa Cruz), and TBP (Abcam) at 4 °C with constant shaking. After incubating with HRP-labeled secondary antibodies (Cell Signaling) at room temperature for 2 h, the protein bands were detected using the Pierce™ ECL Western Blotting Substrate (Thermo Scientific).

Free full text: Click here

Li R., Wang D., Li H., Lei X., Liao W, & Liu X.Y. (2023). Identification of Piezo1 as a potential target for therapy of colon cancer stem-like cells. Discover. Oncology, 14, 95.

Publication 2023

β-actin NFAT1 NFAT2 NFAT3 NFAT4 HRP-labeled secondary antibodies Pierce™ ECL Western Blotting Substrate

Actin Antibodies Protein

Corresponding Organization : Southern University of Science and Technology

Top 5 similar protocols

Variable analysis

independent variables

Primary antibodies targeting β-actin, Piezo1, NFAT1, NFAT2, NFAT3, NFAT4, and TBP

dependent variables

Protein band detection

control variables

Overnight incubation of blots with primary antibodies at 4 °C with constant shaking
Incubation with HRP-labeled secondary antibodies at room temperature for 2 h
Use of Pierce™ ECL Western Blotting Substrate (Thermo Scientific) for protein band detection

controls

No positive or negative controls were explicitly mentioned in the provided information.

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!