CypK-N-Cyclopropene-L-Lysine-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemECypKCat.No.:HY-134669CASNo.:1610703-09-7Synonyms:N-Cyclopropene-L-Lysine分?式:C??H??N?O?分?量:256.3作?靶點(diǎn):Others作?通路:Others儲(chǔ)存?式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY?物活性CypK(N-Cyclopropene-L-Lysine)賴氨酸的環(huán)丙烯衍?物，通過因編碼擴(kuò)增有效地整合到抗體中。CypK?個(gè)最?的?物正交柄，?于創(chuàng)建穩(wěn)定的治療蛋?偶聯(lián)物。體外研究Guidelines(Followingisourrecommendedprotocol.Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds).CypKassay[1](expresstheantibody):1.ThawavialofHEKsuspensioncellsina250mLflaskcontaining50mLofexpressionmediumsupplementedwith100units/mLpenicillin,100μg/mLstreptomycin,and250ng/mLamphotericinB.Keepthecellsat37oCwith8%CO2inhumidifiedincubatorsequippedwithashakerat125rpm.Splitcellsto0.3-0.5x10[6]cells/mL(every2-3days)atleast2timesbeforetransfecting.2.Whenadensityof2.5x10[6]cells/mLisreached(2-3daysaftersplitting),prepareafreshsolutionof100mMCypK.Forthispurpose,weigh64mgofCypK,add2.5mLof0.1sodiumhydroxide,vortex,spindowntorecoverallundissolvedparticlesandsonicate.3.Add2.5mLofCypK(100mMin0.1MNaOH)to42.5mLofexpressionmediumsupplementedwithantibiotics.Mixwell,add250μLof0.1MHCl,andsterilizeusinga0.22μmfilter.4.Dilute50μgofHCandLCpKym1plasmidsto2.5mLwithreducedserummedium.Inaseparatetube,dilute135μLoftransfectionreagentto2.5mLwithreducedserummedium.5.Fiveminutesafterpreparingthesolutions,mixtheplasmidsandthetransfectionreagentsolutionandincubatefor20mintoallowtheformationofcomplexesbetweentheDNAandthetransfectionreagent.6.Inthemeantime,centrifuge125millioncellsatthetargetdensityfor5minat500xg,resuspendwiththeexpressionmediumcontainingCypKandaddtheDNA–transfectionreagentmixture.1/2MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE7.Afterincubatingcellsfor20h,add250μLoftransfectionreagentenhancersincludedinthekit.8.Harvestantibodiesfromthesupernatant6-7daysafteradditionofCypK(nochangeofmediumisrequiredduringexpression).REFERENCES[1].Oller-SalviaB.GeneticEncodingofaNon-CanonicalAminoAcidfortheGenerationofAntibody-DrugConjugatesThroughaFastBioorthogonalReaction.JVisExp.2018Sep14;(139):58066.[2].Oller-SalviaB,et,al.RapidandEfficientGenerationofStableAntibody-DrugConjugatesviaanEncodedCyclopropeneandanInverse-Electron-DemandDiels-AlderReaction.AngewChemIntEdEngl.2018Mar5;57(11):2831-2834.McePdfHeightCaution:Producthasnotbeenfullyvalidatedformedicalappli