Cycloheximide (Naramycin A), an antifungal antibiotic, is an eukaryote protein synthesis inhibitor, with IC₅₀s of 532.5 nM and 2880 nM for protein synthesis and RNA synthesis in vivo, respectively. Cycloheximide suppresses ferroptosis and inhibits autophagy^[1].

IC50: 532.5 nM (protein synthesis), 2.88 μM (RNA synthesis)^[1]

Cycloheximide (CHX) is the most common laboratory reagent used to inhibit protein synthesis. Cycloheximide has been shown to block the elongation phase of eukaryotic translation. Cycloheximide binds the ribosome and inhibits eEF2-mediated translocation. Surprisingly, Cycloheximide allows one complete translocation cycle to proceed before halting any further elongation. Cycloheximide has been speculated that Cycloheximide requires an E-site bound deacylated tRNA for activity^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

The mice receive Cycloheximide injections at 30, 60, or 120 mg/kg prior to training with a 200 µA shock. There is a significant effect of Cycloheximide on latencies on the memory test trials (P<0.001). In saline controls, this shock level results in latencies on the test trial that are significantly higher than those at training. Injections of the lowest dose of Cycloheximide tested, 30 mg/kg, results in latencies on the test trial that are significantly higher than those seen in the saline control group. Mice receiving either of the two higher doses of Cycloheximide has latencies on the test trial that are comparable to those of the saline group, i.e., the higher doses neither enhanced nor impaired memory under these conditions, resulting in an inverted-U dose-response curve for Cycloheximide enhancement of memory^[2]. Infarct volume, as measured by morphometric analysis of infarct areas with triphenyl tetrazolium chloride (TTC), is significantly reduced by 92% and 61% when Cycloheximide is given 0 or 6 hr after HI respectively, but shows an insignificant trend in infarct reduction if Cycloheximide is administered 12 hr after hypoxia-ischemia (HI) compared to the HI control group, and no protective effect is observed when administration is delayed until 24 hr after HI^[3].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

281.35

C₁₅H₂₃NO₄

66-81-9

放线菌酮

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

In Vitro: 

DMSO : 125 mg/mL (444.29 mM; Need ultrasonic)

H₂O : 20 mg/mL (71.09 mM; Need ultrasonic and warming)

*

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (protect from light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.08 mg/mL (7.39 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (7.39 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.08 mg/mL (7.39 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (7.39 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 3.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.08 mg/mL (7.39 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (7.39 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Schneider-Poetsch T, et al. Inhibition of eukaryotic translation elongation by cycloheximide and lactimidomycin. Nat Chem Biol. 2010 Mar;6(3):209-217.

  [2]. Gold PE, et al. Cycloheximide impairs and enhances memory depending on dose and footshock intensity. Behav Brain Res. 2012 Aug 1;233(2):293-7.

  [3]. Park W S, et al. Therapeutic window for cycloheximide treatment after hypoxic-ischemic brain injury in neonatal rats. J Korean Med Sci. 2006 Jun;21(3):490-4.

To test cell proliferation, 3000-5000 cells (HeLa, HTB1 and HEK 293T cells; Jurkat, BT 474, HCC 1395, HCC 1937, HCC 2218 and MDA MB231 cells; MCF 10A) per well are plated in a 96-well plate and allowed to adhere overnight. Cycloheximide dissolved in DMSO at the indicated concentrations (0.1 nM-1000μM) are then added and cells are incubated for a further 24 h. [³H]-thymidine is added at 1 μCi per well and incubation is continued for an additional 7 h. Cells are washed twice with PBS and then trypsinized before they are collected with a Tomtec harvester and bound to GF/C filter mats. Thymidine uptake is then measured by scintillation counting^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[2]
Male ICR mice (approximately 2 months old) are used in this experiment. Cycloheximide is administered IP at concentrations of 0 (saline controls), 30, 60, or 120 mg/kg. Cycloheximide injections are administered 30 min prior to training. The 120 mg/kg dose is commonly used to study amnesia in mice. Note that amnestic Cycloheximide doses are much lower in rats (1-3 mg/kg) than in mice, consistent with a similar difference in LD50s for rats and mice. Cycloheximide doses of 120-150 mg/kg result in approximately 95% inhibition of brain protein synthesis as measured 30-60 min after injection; the dose of 30 mg/kg produces approximately 80% inhibition of brain protein synthesis.
Rats^[3]
Unilateral carotid artery ligation is performed in 7-day old Sprague Dawley rat pups under methoxyflurane anesthesia. The neck is incised in the midline, and the right common carotid artery is permanently ligated with 4-0 silk. Total time of surgery in each animal never exceeded 5 min. Following surgery, rats are returned to their mother for recovery and feeding for 2 hr. The pups are then exposed to a 100 min-period of hypoxia (8% O₂, 92% N₂) by placing them in an airtight chamber partially submerged in a temperature controlled water bath to maintain the ambient temperature inside the chamber at a constant 36°C. In the HI with Cycloheximide treatment group, the rat pups receive an intraperitoneal injection of Cycloheximide at a dose of 0.6 mg/kg at 0, 6, 12 or 24 hr of recovery, and an equal volume of normal saline is given to a HI control group. Then, the rat pups are returned to their dam until sacrifice; the whole brain tissue is obtained under deep pentobarbital anesthesia (60 mg/kg, intraperitoneal) for flow cytometry and triphenyl tetrazolium chloride (TTC) at 48 and 72 hr after HI, respectively.

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Schneider-Poetsch T, et al. Inhibition of eukaryotic translation elongation by cycloheximide and lactimidomycin. Nat Chem Biol. 2010 Mar;6(3):209-217.

  [2]. Gold PE, et al. Cycloheximide impairs and enhances memory depending on dose and footshock intensity. Behav Brain Res. 2012 Aug 1;233(2):293-7.

  [3]. Park W S, et al. Therapeutic window for cycloheximide treatment after hypoxic-ischemic brain injury in neonatal rats. J Korean Med Sci. 2006 Jun;21(3):490-4.