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Cycloheximide is a protein synthesis inhibitor in eukaryotes

Although the protein synthesis inhibitor cycloheximide ..

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Cycloheximide|Antibiotic,inhibiter of protein synthesis …

N2 - Sanguinarine, a benzophenanthridine alkaloid, has anticancer potential through induction of cell death. We previously demonstrated that sanguinarine treatment at a low concentration (1.5 μg/ml) induced apoptosis in K562 human erythroleukemia cells, and a high concentration (12.5 μg/ml) induced the morphology of blister formation or oncosis-blister cell death (BCD). Treatment of cells at an intermediate sanguinarine concentration (6.25 μg/ml) induced diffuse swelling or oncosis-diffuse cell swelling (DCS). To assess the underlying mechanism of sanguinarine-induced apoptosis and oncosis-BCD in K562 cells, we studied their response to pre-treatment with two chemical compounds: aurintricarboxylic acid (ATA) and cycloheximide (CHX). The pretreatment effects of both chemical compounds on apoptosis and oncosis-BCD were evaluated by measuring multiple parameters using quantitative morphology, electron microscopy, terminal deoxynucleotidyl transferase (TdT) end-labeling and annexin-V-binding. ATA, a DNA endonuclease inhibitor, efficiently prevented DNA nicking and inhibited apoptosis almost completely and oncosis-BCD by about 40%, while CHX, a protein synthesis inhibitor, failed to inhibit both apoptosis and oncosis-BCD. These results demonstrate, first, the importance of endonuclease in sanguinarine-induced apoptosis and to some extent in oncosis-BCD and, second, that this inhibition does not require de novo protein synthesis.

Inhibition of protein synthesis in vitro by cycloheximide and related glutarimide antibiotics

I need an advice. I plan to treat MCF-7 cells with my test compound. First I need to stop nascent protein synthesis because I want to work only with proteins existing before treatment. For this purpose I want to use cycloheximide but I don't know what concentration I should use and how long incubate with cycloheximide before adding my test compound. Should I wash cells from medium containing cycloheximide before adding my compound? Maybe you recommend different method than this with cycloheximide. Thank you in advance for some information.

Inhibition of Mammalian Mitochondrial Protein Synthesis …

For understanding the mechanisms of consolidation, it is important to know how protein synthesis inhibitors affect hippocampal neurons.

Sanguinarine, a benzophenanthridine alkaloid, has anticancer potential through induction of cell death. We previously demonstrated that sanguinarine treatment at a low concentration (1.5 μg/ml) induced apoptosis in K562 human erythroleukemia cells, and a high concentration (12.5 μg/ml) induced the morphology of blister formation or oncosis-blister cell death (BCD). Treatment of cells at an intermediate sanguinarine concentration (6.25 μg/ml) induced diffuse swelling or oncosis-diffuse cell swelling (DCS). To assess the underlying mechanism of sanguinarine-induced apoptosis and oncosis-BCD in K562 cells, we studied their response to pre-treatment with two chemical compounds: aurintricarboxylic acid (ATA) and cycloheximide (CHX). The pretreatment effects of both chemical compounds on apoptosis and oncosis-BCD were evaluated by measuring multiple parameters using quantitative morphology, electron microscopy, terminal deoxynucleotidyl transferase (TdT) end-labeling and annexin-V-binding. ATA, a DNA endonuclease inhibitor, efficiently prevented DNA nicking and inhibited apoptosis almost completely and oncosis-BCD by about 40%, while CHX, a protein synthesis inhibitor, failed to inhibit both apoptosis and oncosis-BCD. These results demonstrate, first, the importance of endonuclease in sanguinarine-induced apoptosis and to some extent in oncosis-BCD and, second, that this inhibition does not require de novo protein synthesis.

50-100 micromolar cycloheximide should be heaps, it is apparently effective within 15 minutes of addition, but it would probably be best if you waited for 30 min to one hour to ensure effectiveness. As far as I recall, CHX only stops about 70-80% of protein synthesis, and emetine is more effective, though CHX is more commonly used. Ideally you will do titrations of the amount and times to use CHX before deciding on what you will do for your experiments.

AS to whether you should wash your cells after removal of CHX, yes, you don't want residue interfering with your experiment.

Cycloheximide is a protein synthesis inhibitor in ..

Effect of the protein synthesis inhibitor cycloheximide on RNA synthesis ..

AB - Sanguinarine, a benzophenanthridine alkaloid, has anticancer potential through induction of cell death. We previously demonstrated that sanguinarine treatment at a low concentration (1.5 μg/ml) induced apoptosis in K562 human erythroleukemia cells, and a high concentration (12.5 μg/ml) induced the morphology of blister formation or oncosis-blister cell death (BCD). Treatment of cells at an intermediate sanguinarine concentration (6.25 μg/ml) induced diffuse swelling or oncosis-diffuse cell swelling (DCS). To assess the underlying mechanism of sanguinarine-induced apoptosis and oncosis-BCD in K562 cells, we studied their response to pre-treatment with two chemical compounds: aurintricarboxylic acid (ATA) and cycloheximide (CHX). The pretreatment effects of both chemical compounds on apoptosis and oncosis-BCD were evaluated by measuring multiple parameters using quantitative morphology, electron microscopy, terminal deoxynucleotidyl transferase (TdT) end-labeling and annexin-V-binding. ATA, a DNA endonuclease inhibitor, efficiently prevented DNA nicking and inhibited apoptosis almost completely and oncosis-BCD by about 40%, while CHX, a protein synthesis inhibitor, failed to inhibit both apoptosis and oncosis-BCD. These results demonstrate, first, the importance of endonuclease in sanguinarine-induced apoptosis and to some extent in oncosis-BCD and, second, that this inhibition does not require de novo protein synthesis.

The rates of accumulation of newly synthesized catecholamines and endogenous catecholamine levels in mice were determined after treatment with cycloheximide, acetoxycycloheximde, puromycin, and anisomycin. The rates of accumulation were found to be decreased by all antibiotics tested, weakening the assumption that their amnestic effects are due solely to inhibition of protein synthesis.

The ability of cycloheximide to inhibit protein synthesis was not affected by ..
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