Home>>Signaling Pathways>> Ubiquitination/ Proteasome>> Autophagy>>Cycloheximide


Catalog No.: GC17198

Cycloheximide is an antibiotic that inhibits protein synthesis at the translation level

Cycloheximide Chemical Structure

Size Price Stock Qty
10mM (in 1mL DMSO)
In stock
In stock
In stock
In stock
In stock

Customer Reviews

Based on customer reviews.

Tel: (626) 353-8530 Email: sales@glpbio.com

Sample solution is provided at 25 µL, 10mM.

Product Citations

Product Documents

Quality Control & SDS

View current batch:


Cell experiment [1]:

Cell lines


Preparation Method

Hepatocyte suspensions(2 x 106 cells/ml) were incubated in polycarbonate flasks, at 37℃, under a constant stream of 95% O2- 5% CO2, with constant shaking (72 cycles/min), for 15 min before addition of cycloheximide.

Reaction Conditions

Hepatocytes were treated with a range of concentrations of cycloheximide from1 x 10-7 to 5 x 10-3 M).


Low, non-toxic dose of cycloheximide provides reasonable assurance that protein synthetic ability could be perturbed without causing undue alterations in other biochemical functions of the cell. A nontoxic dose of cycloheximide (1 μM) inhibited termination to a greater extent than other translational steps. This effect showed a dose-dependent manner.

Animal experiment [2]:

Animal models

Male Sprague-Dawley rats,weighing between 160-230 g

Preparation Method

Mice were maintained on Purina Chow and water ad libitum. Food was withdrawn from the animals the night before they were given 14C-ANIT, Cycloheximide was injected 1/2, 1, 2, 4, 8 or 24 h before ANIT administration.

Dosage form

2 mg/kg


Cycloheximide is capable of protecting against ANIT-induced hyperbilirubinemia even if given 24 h before ANIT. It became apparent that cycloheximide treatment resulted in substantially reduced amounts of ANIT-equivalents in all tissues examined, even if cycloheximide was given 24 h before ANIT.


[1]. Helinek TG, et al. Initial inhibition and recovery of protein synthesis in cycloheximide-treated hepatocytes. Biochem Pharmacol. 1982 Apr 1;31(7):1219-25.

[2]. Lock S, et al. Effect of cycloheximide on the distribution of alpha -naphthylisothiocyanate in rats. Exp Mol Pathol. 1974 Oct;21(2):237-45.


Cycloheximide is an antibiotic that inhibits protein synthesis at the translation level, acting exclusively on cytoplasmic (80s) ribosomes of eukaryotes. Cycloheximide affected all the energy-dependent stages in the protein-synthesizing process. However, the initiation seems the most sensitive. Cycloheximide also affects respiration, ion uptake, amino acid biosynthesis, and DNA and RNA synthesis, effects that are probably secondary to its effect on protein synthesis.[1]

In vitro study indicated that Cycloheximide at 1 μM inhibited [3H]leucine incorporation into both cellular and secreted proteins by at least 86%, without having deleterious effects on membrane integrity as indicated by trypan blue uptake and lactate dehydrogenase release. Larger size class polysomes (7+) were increased by Cycloheximide treatment and remained increased during recovery. [2]

In vivo analysis indicated that Cycloheximide produced initial hyperactivity. This initial hyperactivity was apparent within 3 minutes after injection of the Cycloheximide. Cycloheximide affects activity by acting on the brain, and this is unrelated to its inhibition of protein synthesis. In addition, Cycloheximide’s effects on activity did not appear to be responsible for its amnesic action. However, Cycloheximide might have some other property, unrelated to inhibition of cerebral protein synthesis, that is responsible for its amnesic effect.[3]

[1]. Marcos R, et al. Effect of Cycloheximide on different stages of Drosophila melanogaster. Toxicol Lett. 1982 Sep;13(1-2):105-12.
[2].Helinek TG, et al. Initial inhibition and recovery of protein synthesis in cycloheximide-treated hepatocytes. Biochem Pharmacol. 1982 Apr 1;31(7):1219-25.
[3]. Segal DS, et al. Cycloheximide: its effects on activity are dissociable from its effects on memory. Science. 1971 Apr 2;172(3978):82-4.

Chemical Properties

Cas No. 66-81-9 SDF
Synonyms Naramycin A; Actidione; 3-[2-(3,5-Dimethyl-2-oxocyclohexyl)-2-hydroxyethyl]glutarimide
Chemical Name 4-[(2R)-2-[(1S,3S,5S)-3,5-dimethyl-2-oxocyclohexyl]-2-hydroxyethyl]piperidine-2,6-dione
Canonical SMILES CC1CC(C(=O)C(C1)C(CC2CC(=O)NC(=O)C2)O)C
Formula C15H23NO4 M.Wt 281.4
Solubility ≥ 14.07 mg/mL in DMSO, ≥ 57.6 mg/mL in EtOH, ≥ 14.05 mg/mL in Water with ultrasonic and warming Storage 4°C, protect from light
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.
Shipping Condition Evaluation sample solution : ship with blue ice
All other available size: ship with RT , or blue ice upon request

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.

  • Molarity Calculator

  • Dilution Calculator

**When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and MSDS / CoA (available online).


Research Update

Cycloheximide Chase Analysis of Protein Degradation in Saccharomyces cerevisiae

J Vis Exp2016 Apr 18;(110):53975.PMID: 27167179DOI: 10.3791/53975

Regulation of protein abundance is crucial to virtually every cellular process. Protein abundance reflects the integration of the rates of protein synthesis and protein degradation. Many assays reporting on protein abundance (e.g., single-time point western blotting, flow cytometry, fluorescence microscopy, or growth-based reporter assays) do not allow discrimination of the relative effects of translation and proteolysis on protein levels. This article describes the use of cycloheximide chase followed by western blotting to specifically analyze protein degradation in the model unicellular eukaryote, Saccharomyces cerevisiae (budding yeast). In this procedure, yeast cells are incubated in the presence of the translational inhibitor cycloheximide. Aliquots of cells are collected immediately after and at specific time points following addition of cycloheximide. Cells are lysed, and the lysates are separated by polyacrylamide gel electrophoresis for western blot analysis of protein abundance at each time point. The cycloheximide chase procedure permits visualization of the degradation kinetics of the steady state population of a variety of cellular proteins. The procedure may be used to investigate the genetic requirements for and environmental influences on protein degradation.

Discovery of C13-Aminobenzoyl Cycloheximide Derivatives that Potently Inhibit Translation Elongation

J Am Chem Soc2021 Sep 1;143(34):13473-13477.PMID: 34403584DOI: 10.1021/jacs.1c05146

Employed for over half a century to study protein synthesis, cycloheximide (CHX, 1) is a small molecule natural product that reversibly inhibits translation elongation. More recently, CHX has been applied to ribosome profiling, a method for mapping ribosome positions on mRNA genome-wide. Despite CHX's extensive use, CHX treatment often results in incomplete translation inhibition due to its rapid reversibility, prompting the need for improved reagents. Here, we report the concise synthesis of C13-amide-functionalized CHX derivatives with increased potencies toward protein synthesis inhibition. Cryogenic electron microscopy (cryo-EM) revealed that C13-aminobenzoyl CHX (8) occupies the same site as CHX, competing with the 3' end of E-site tRNA. We demonstrate that 8 is superior to CHX for ribosome profiling experiments, enabling more effective capture of ribosome conformations through sustained stabilization of polysomes. Our studies identify powerful chemical reagents to study protein synthesis and reveal the molecular basis of their enhanced potency.

Validity of cycloheximide chylomicron flow blocking method for the evaluation of lymphatic transport of drugs

Br J Pharmacol2021 Dec;178(23):4663-4674.PMID: 34365639DOI: 10.1111/bph.15644

Background and purpose: Lymphatic transport of drugs after oral administration is an important mechanism for absorption of highly lipophilic compounds. Direct measurement in lymph duct cannulated animals is the gold standard method, but non-invasive cycloheximide chylomicron flow blocking method has gained popularity recently. However, concerns about its reliability have been raised. The aim of this work was to investigate the validity of cycloheximide chylomicron flow blocking method for the evaluation of lymphatic transport using model compounds with high to very high lipophilicity, that is, abiraterone and cinacalcet.
Experimental approach: Series of pharmacokinetic studies were conducted with abiraterone acetate and cinacalcet hydrochloride after enteral/intravenous administration to intact, lymph duct cannulated and/or cycloheximide pre-treated rats.
Key results: Mean total absolute oral bioavailability of abiraterone and cinacalcet was 7.0% and 28.7%, respectively. There was a large and significant overestimation of the lymphatic transport extent by the cycloheximide method. Mean relative lymphatic bioavailability of abiraterone and cinacalcet in cycloheximide method was 28-fold and 3-fold higher than in cannulation method, respectively.
Conclusion and implications: Cycloheximide chylomicron flow blocking method did not provide reliable results on lymphatic absorption and substantially overestimated lymphatic transport for both molecules, that is, abiraterone and cinacalcet. This non-invasive method should not be used for the assessment of lymphatic transport and previously obtained data should be critically revised.

Proteome-wide mapping of short-lived proteins in human cells

Mol Cell2021 Nov 18;81(22):4722-4735.e5.PMID: 34626566DOI: 10.1016/j.molcel.2021.09.015

Rapid protein degradation enables cells to quickly modulate protein abundance. Dysregulation of short-lived proteins plays essential roles in disease pathogenesis. A focused map of short-lived proteins remains understudied. Cycloheximide, a translational inhibitor, is widely used in targeted studies to measure degradation kinetics for short-lived proteins. Here, we combined cycloheximide chase assays with advanced quantitative proteomics to map short-lived proteins under translational inhibition in four human cell lines. Among 11,747 quantified proteins, we identified 1,017 short-lived proteins (half-lives ≤ 8 h). These short-lived proteins are less abundant, evolutionarily younger, and less thermally stable than other proteins. We quantified 103 proteins with different stabilities among cell lines. We showed that U2OS and HCT116 cells express truncated forms of ATRX and GMDS, respectively, which have lower stability than their full-length counterparts. This study provides a large-scale resource of human short-lived proteins under translational arrest, leading to untapped avenues of protein regulation for therapeutic interventions.

Cycloheximide in dermatology

Acta Derm Venereol1997 May;77(3):240.PMID: 9188885DOI: 10.2340/0001555577240

Metabolism in aquatic ectotherms evaluated by oxygen consumption rates reflects energetic costs including those associated with protein synthesis. Metabolism is influenced by nutritional status governed by feeding, nutrient intake and quality, and time without food. However, little is understood about contribution of protein synthesis to crustacean energy metabolism. This study is the first using a protein synthesis inhibitor cycloheximide to research contribution of cycloheximide-sensitive protein synthesis to decapod crustacean metabolism. Juvenile Sagmariasus verreauxi were subject to five treatments: 2-day fasted lobsters sham injected with saline; 2-day fasted lobsters injected with cycloheximide; 10-day starved lobsters injected with cycloheximide; post-prandial lobsters fed with squid Nototodarus sloanii with no further treatment; and post-prandial lobsters injected with cycloheximide. Standard and routine metabolic rates in starved lobsters were reduced by 32% and 41%, respectively, compared to fasted lobsters, demonstrating metabolic downregulation with starvation. Oxygen consumption rates of fasted and starved lobsters following cycloheximide injection were reduced by 29% and 13%, respectively, demonstrating protein synthesis represents only a minor component of energy metabolism in unfed lobsters. Oxygen consumption rate of fed lobsters was reduced by 96% following cycloheximide injection, demonstrating protein synthesis in decapods contributes a major proportion of specific dynamic action (SDA). SDA in decapods is predominantly a post-absorptive process likely related to somatic growth. This work extends previously limited knowledge on contribution of protein synthesis to crustacean metabolism, which is crucial to explore the relationship between nutritional status and diet quality and how this will affect growth potential in aquaculture species.


Review for Cycloheximide

Average Rating: 5 ★★★★★ (Based on Reviews and 36 reference(s) in Google Scholar.)

5 Star
4 Star
3 Star
2 Star
1 Star
Review for Cycloheximide

GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.

Required fields are marked with *

You may receive emails regarding this submission. Any emails will include the ability to opt-out of future communications.