Search

2 users have reported that they have successfully carried out the experiment using this protocol.
Analysis of Protein Stability by the Cycloheximide Chase Assay   

Download PDF How to cite Favorites 1 Q&A Share your feedback Cited by

In this protocol

Ask a question to the author

Original research article

A brief version of this protocol appeared in:
Oncogene
Jun 2014

Abstract

Comparison of protein stability in eukaryotic cells has been achieved by cycloheximide, which is an inhibitor of protein biosynthesis due to its prevention in translational elongation. It is broadly used in cell biology in terms of determining the half-life of a given protein and has gained much popularity in cancer research. Here we present a full cycloheximide chase assay in our laboratory using a lung adenocarcinoma cell line, CL1-5, as a model.

Copyright: © 2015 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Kao, S., Wang, W., Chen, C., Chang, Y., Wu, Y., Wang, Y., Wang, S., Nesvizhskii, A. I., Chen, Y., Hong, T. and Yang, P. (2015). Analysis of Protein Stability by the Cycloheximide Chase Assay. Bio-protocol 5(1): e1374. DOI: 10.21769/BioProtoc.1374.
Q&A

Please login to post your questions/comments. Your questions will be directed to the authors of the protocol. The authors will be requested to answer your questions at their earliest convenience. Once your questions are answered, you will be informed using the email address that you register with bio-protocol.
You are highly recommended to post your data including images for the troubleshooting.

You are highly recommended to post your data (images or even videos) for the troubleshooting. For uploading videos, you may need a Google account because Bio-protocol uses YouTube to host videos.

Tao Zhang
Institute of Neuroscience, CAS
Hello,I have two questions. Do I need to mixing the lysate adequately before transfer it to -80? How could I determine the concentration of CHX when I use the cells of rat embryonic fibroblast. Should I treated the cells with CHX with different concentrations for 8 hours and observe the apoptosis markers and some protein with well-known half-life? Thank you very much.
9/2/2015 1:08:52 AM Reply