How does cycloheximide chase work?
Cycloheximide chase assays are an experimental technique used in molecular and cellular biology to measure steady state protein stability. Cycloheximide is a drug that inhibits the elongation step in eukaryotic protein translation, thereby preventing protein synthesis.
Is cycloheximide reversible?
Here we show that cycloheximide is nontoxic and its effect is reversible, allowing protein synthesis to reinitiate.
What is the purpose of the cycloheximide in the plates?
Cycloheximide is added to some media (mainly solid media), such as YPD agar, WLN agar and MRS-AJ agar, to suppress the growth of most yeast.
What does cycloheximide treat?
It is used as a rodenticide and other animal pesticide. It is also used in media to detect unwanted bacteria in beer fermentation by suppressing yeasts and molds growth in test medium. The translational elongation freezing properties of cycloheximide are also used for ribosome profiling / translational profiling.
Is cycloheximide an antibiotic?
Cycloheximide is a dicarboximide that is 4-(2-hydroxyethyl)piperidine-2,6-dione in which one of the hydrogens attached to the carbon bearing the hydroxy group is replaced by a 3,5-dimethyl-2-oxocyclohexyl group. It is an antibiotic produced by the bacterium Streptomyces griseus.
Is cycloheximide an antifungal?
Cycloheximide is an antibiotic with significant antifungal properties. It is produced by some streptomycin-producing strains of Streptomyces griseus and acts by inhibiting protein synthesis.
How do you use cycloheximide Chase?
Cycloheximide chase may be implemented to analyze the degradation of a variety of yeast proteins and may be adapted to study protein stability in other eukaryotic cells. As described in this protocol, cycloheximide treatment is followed by cell lysis and detection of protein abundance by western blot analysis.
Does cycloheximide affect protein stability in eukaryotic cells?
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.
How do you measure protein abundance after Cycloheximide treatment?
As described in this protocol, cycloheximide treatment is followed by cell lysis and detection of protein abundance by western blot analysis. Depending on the application, however, protein abundance following cycloheximide treatment may be assessed by a range of techniques, as appropriate for research objectives.
Is there a full cycloheximide Chase assay for lung adenocarcinoma?
Here we present a full cycloheximide chase assay in our laboratory using a lung adenocarcinoma cell line, CL1-5, as a model. Materials and Reagents