Measuring Myotube Protein Synthesis

Myotubes were grown and treated as described previously. To determine the rate of protein synthesis we utilized SUnSET methodology, as described (10 (link), 11 (link)). Briefly, puromycin (Sigma-Aldrich) was administered to the media at a final concentration of 1 μM exactly 30 min before cells were collected in ice-cold homogenizing buffer. Anti-puromycin was purchased from Millipore (Kilsyth, Victoria, Australia) and immunoblotting was used to detect changes in puromycin incorporation as described (9 (link), 10 (link)).

Free full text: Click here

Caldow M.K., Ham D.J., Trieu J., Chung J.D., Lynch G.S, & Koopman R. (2019). Glycine Protects Muscle Cells From Wasting in vitro via mTORC1 Signaling. Frontiers in Nutrition, 6, 172.

Publication 2019

puromycin Anti-puromycin

Buffer Cells Cold Myotubes Protein synthesis Puromycin

Corresponding Organization : University of Melbourne

Top 5 similar protocols

Variable analysis

independent variables

Puromycin (Sigma-Aldrich) administered to the media at a final concentration of 1 μM

dependent variables

Changes in puromycin incorporation

control variables

Time of puromycin administration (exactly 30 min before cells were collected)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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!