Hi,
I am treating my cells with a RNA Pol inhibitor which inhibits transcription of rDNA. Ribosome production will be inhibited and I would like to show how this affects the rate of protein synthesis?
Can you suggest and assays I could use to investigate this please?
Many thanks in advance
The rate of protein synthesis can be measured as the rate of incorporation of radiolabeled amino acids into acid-precipitable counts.
Thank you Adam, do you know of any standard assays I could use or any radiolabeled amino acids which are preferred to measure the rate of protein synthesis?
Your best bet would be to find some publications in which this technique was used for the purpose in the same type of cells you are planning to use. You would want to use an amino acid that is well-represented in proteins, such as leucine or valine. Use 3H because of its high specific radioactivity and safety.
See page 11 of this paper for an example of the method for measuring protein synthesis in cultured cells by 3H-Leu incorporation and TCA precipitation.
http://www.cell.com/immunity/pdf/S1074-7613(03)00051-7.pdf
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