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Medchemexpress/Cycloheximide(Synonyms: Naramycin A; Actidione)/HY-12320/10mM*1mL in DMSO
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Medchemexpress/Cycloheximide(Synonyms: Naramycin A; Actidione)/HY-12320/10mM*1mL in DMSO
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Cycloheximideisaninhibitorofproteinbiosynthesisineukaryoticorganisms,withIC50of532.5nMand2880nMforproteinsynthesisandRNAsynthesisinvivo,respectively.

CustomerValidation

  • PLoSPathog.2017Jun19;13(6):e1006449.
  • CellDeathDis.2017Sep14;8(9):e3048.
  • BiochimBiophysActa.2016Dec;1859(12):1527-1537.
  • Oncotarget.2016May10;7(19):27176-84.
  • BiomedResInt.2016;2016:8145495.
  • BiochemBiophysResCommun.2017Sep2;490(4):1168-1175.
Description

Cycloheximideisaninhibitorofproteinbiosynthesisineukaryoticorganisms,withIC50of532.5nMand2880nMforproteinsynthesisandRNAsynthesisinvivo,respectively.

IC50&Target

IC50:532.5nM(proteinsynthesis),2.88μM(RNAsynthesis)[1]

InVitro

Cycloheximide(CHX)isthemostcommonlaboratoryreagentusedtoinhibitproteinsynthesis.Cycloheximidehasbeenshowntoblocktheelongationphaseofeukaryotictranslation.CycloheximidebindstheribosomeandinhibitseEF2-mediatedtranslocation.Surprisingly,Cycloheximideallowsonecompletetranslocationcycletoproceedbeforehaltinganyfurtherelongation.CycloheximidehasbeenspeculatedthatCycloheximiderequiresanE-sitebounddeacylatedtRNAforactivity[1].

InVivo

ThemicereceiveCycloheximideinjectionsat30,60,or120mg/kgpriortotrainingwitha200µAshock.ThereisasignificanteffectofCycloheximideonlatenciesonthememorytesttrials(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="">[2].Infarctvolume,asmeasuredbymorphometricanalysisofinfarctareaswithtriphenyltetrazoliumchloride(TTC),issignificantlyreducedby92%and61%whenCycloheximideisgiven0or6hrafterHIrespectively,butshowsaninsignificanttrendininfarctreductionifCycloheximideisadmiNISTered12hrafterhypoxia-ischemia(HI)comparedtotheHIcontrolgroup,andnoprotectiveeffectisobservedwhenadministrationisdelayeduntil24hrafterHI[3].

References
  • [1].Schneider-PoetschT,etal.Inhibitionofeukaryotictranslationelongationbycycloheximideandlactimidomycin.NatChemBiol.2010Mar;6(3):209-217.

    [2].GoldPE,etal.Cycloheximideimpairsandenhancesmemorydependingondoseandfootshockintensity.BehavBrainRes.2012Aug1;233(2):293-7.

    [3].ParkWS,etal.Therapeuticwindowforcycloheximidetreatmentafterhypoxic-ischemicbraininjuryinneonatalrats.JKoreanMedSci.2006Jun;21(3):490-4.

PreparingStockSolutions
ConcentrationVolumeMass1mg5mg10mg
1mM3.5543mL17.7715mL35.5429mL
5mM0.7109mL3.5543mL7.1086mL
10mM0.3554mL1.7771mL3.5543mL
Pleaserefertothesolubilityinformationtoselecttheappropriatesolvent.
CellAssay
[1]

Cycloheximideisdissolvedinstocksolutions,andthenseriallydilutedwithappropriatemediabeforeuse[1].

Totestcellproliferation,3000-5000cells(HeLa,HTB1andHEK293Tcells;Jurkat,BT474,HCC1395,HCC1937,HCC2218andMDAMB231cells;MCF10A)perwellareplatedina96-wellplateandallowedtoadhereovernight.CycloheximidedissolvedinDMSOattheindicatedconcentrations(0.1nM-1000μM)arethenaddedandcellsareincubatedforafurther24h.[3H]-thymidineisaddedat1μCiperwellandincubationiscontinuedforanadditional7h.CellsarewashedtwicewithPBSandthentrypsinizedbeforetheyarecollectedwithaTomtecharvesterandboundtoGF/Cfiltermats.Thymidineuptakeisthenmeasuredbyscintillationcounting[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

AnimalAdministration
[2][3]

Cycloheximide(CHX)isdissolvedinsaline(Mice)[2].

Mice[2]
MaleICRmice(approximately2monthsold)areusedinthisexperiment.CycloheximideisadministeredIPatconcentrationsof0(salinecontrols),30,60,or120mg/kg.Cycloheximideinjectionsareadministered30minpriortotraining.The120mg/kgdoseiscommonlyusedtostudyamnesiainmice.NotethatamnesticCycloheximidedosesaremuchlowerinrats(1-3mg/kg)thaninmice,consistentwithasimilardifferenceinLD50sforratsandmice.Cycloheximidedosesof120-150mg/kgresultinapproximately95%inhibitionofbrainproteinsynthesisasmeasured30-60minafterinjection;thedoseof30mg/kgproducesapproximately80%inhibitionofbrainproteinsynthesis.
Rat[3]
Unilateralcarotidarteryligationisperformedin7-dayoldSpragueDawleyratpupsundermethoxyfluraneanesthesia.Theneckisincisedinthemidline,andtherightcommoncarotidarteryispermanentlyligatedwith4-0silk.Totaltimeofsurgeryineachanimalneverexceeded5min.Followingsurgery,ratsarereturnedtotheirmotherforrecoveryandfeedingfor2hr.Thepupsarethenexposedtoa100min-periodofhypoxia(8%O2,92%N2)byplacingtheminanairtightchamberpartiallysubmergedinatemperaturecontrolledwaterbathtomaintaintheambienttemperatureinsidethechamberataconstant36℃.IntheHIwithCycloheximidetreatmentgroup,theratpupsreceiveanintraperitonealinjectionofCycloheximideatadoseof0.6mg/kgat0,6,12or24hrofrecovery,andanequalvolumeofnormalsalineisgiventoaHIcontrolgroup.Then,theratpupsarereturnedtotheirdamuntilsacrifice;thewholebraintissueisobtainedunderdeeppentobarbitalanesthesia(60mg/kg,intraperitoneal)forflowcytometryandtriphenyltetrazoliumchloride(TTC)at48and72hrafterHI,respectively.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

References
  • [1].Schneider-PoetschT,etal.Inhibitionofeukaryotictranslationelongationbycycloheximideandlactimidomycin.NatChemBiol.2010Mar;6(3):209-217.

    [2].GoldPE,etal.Cycloheximideimpairsandenhancesmemorydependingondoseandfootshockintensity.BehavBrainRes.2012Aug1;233(2):293-7.

    [3].ParkWS,etal.Therapeuticwindowforcycloheximidetreatmentafterhypoxic-ischemicbraininjuryinneonatalrats.JKoreanMedSci.2006Jun;21(3):490-4.

MolecularWeight

281.35

Formula

C₁₅H₂₃NO₄

CASNo.

66-81-9

Storage
Powder-20°C3years
 4°C2years
Insolvent-80°C6months
 -20°C1month
Shipping

RoomtemperatureincontinentalUS;mayvaryelsewhere

Solvent&Solubility

DMSO:≥33mg/mL;H2O:20mg/mL(Needultrasonicandwarming)

*"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">

Purity:99.17%