MCE 重组蛋白 冻干粉蛋白溶解方法

www.MedChemE重组蛋白RecombinantProteins高纯度

高生物活性

稳定的一致性

GMP

级蛋白MHCVirusIgGEGFIL-6FcγRIIPD-1EGFRPD-L1IL-6RRASCARRAF-1ERK2STAT3MEK1AKT1Elk1DNAwww.MedChemEInhibitors • ScreeningLibraries • ProteinsMCE重组蛋白MCE重组蛋白产品种类丰富，涵盖细胞因子、病毒蛋白、免疫检查点蛋白、CAR-T蛋白、CD抗原、受体蛋白、生物素标记蛋白、GMP级蛋白、酶等多种分类，可用于人、病毒、小鼠、大鼠、猴、猪、狗、牛、羊、兔、猫、非洲爪蟾等多个物种不同研究领域的生物学研究，包括蛋白结构研究、免疫检测试剂、重组蛋白药物、诊断试剂开发、抗体药物靶点、CAR-T细胞治疗靶点、Fc受体、流感病毒蛋白和细胞因子等热门研究领域。MCE因高品质的产品和优质的售前售后服务，成为全球数百万计的科学家和技术人员值得信赖的实验伙伴。产品种类丰富： 细胞因子和生长因子、免疫检查点蛋白、CAR-T相关蛋白、CD抗原、受体蛋白、酶&调节子、补体系统蛋白、病毒类蛋白、生物素标记蛋白、荧光标记蛋白、GMP级蛋白,泛素相关蛋白等产品物种多样性：人、病毒、小鼠、大鼠、猴、猪、狗、牛、羊、兔、猫、非洲爪蟾、细菌、真菌等低内毒素水平： LAL法验证，内毒素含量低于绝大多数供应标准高纯度： SDS和HPLC检测验证高生物活性： 相关体内/体外生物学实验测定稳定的一致性： 多批次检测数据确认规格齐全： 满足客户的多种预分装规格高纯度ThepurityofhumanBMP-2isgreaterthan95%asanalyzedbySDSunderreducing(R)andnon-reducing(N)condition.kDa M R 120806040302010卓越的批间稳定性TheED50ofMCEhumanVEGF165fromthreedifferentLotsaresimilar.1.0Lot1nm)0.8Lot20.6Lot3(4900.4OD0.2Net0.01010101010VEGF165,Human(ng/mL)

高活性HumanGM-CSFstimulatescellproliferationofTF-1cellswithanED50oflessthan0.5ng/mL.2.0(490nm)1.51.0NetOD0.50.010101010101010GM-CSF,Human(ng/mL)竞品对比TheED50ofhumanVEGF165fromMCE’seachLotislowerthanofCompetitorP.1.0BrandPnm)0.8Lot20.6Lot3(4900.4OD0.2Net0.010-210-1101010VEGF165,Human(ng/mL)www.MedChemEInhibitors • ScreeningLibraries • Proteins冻干粉蛋白溶解方法开盖之前需离心。冻干粉在运输过程中，会因为外力因素粘附于管壁或者管盖上，在开盖之前，先通过离心操作，将冻干粉收集于管底，避免损失和浪费。建议10,000-12,000rpm高速离心30s；若没有高速离心机，3,000-3,500rpm离心5min。离心之后，向冻干粉中加入提供的复溶缓冲液，用枪头轻轻吹打混匀，重悬至浓度不低于100μg/mL。溶解过程中，不可用涡旋震荡仪快速震荡，否则会破坏蛋白的空间结构，进而影响蛋白活性。用复溶缓冲液直接溶解的重组蛋白液体可在2-8°C保存1周。对于周期较短的实验(不超过7天)，可以直接取该条件下保存的重组蛋白溶液加入到培养体系内即可，于一周之内用完；如果实验条件设置的使用浓度低于复溶之后的溶液浓度，请用含载体蛋白的缓冲液进一步稀释，如果稀释液不含载体蛋白，容易导致重组蛋白粘附在管壁或者瓶壁上，使其在溶液中的浓度下降，进一步导致重组蛋白的活性减弱。若要长期保存，则需用含载体蛋白(0.1%BSA，5%HSA，10%FBS，或5%海藻糖)的溶液进一步稀释，然后分装冻存于-20°C至-80°C。复溶后的产品不建议直接冻存于-20°C至-80°C。因为塑料管壁对蛋白有很强的吸附作用，会导致重组蛋白粘附在管壁不易分离，进而导致溶液中重组蛋白的实际浓度偏低，最终导致其活性下降。而载体蛋白可以预先封闭塑料管壁上蛋白结合位点，使重组蛋白不会粘附于管壁。因此长期保存时，在分装冻存前，建议用含载体蛋白的溶液进一步稀释(浓度不低于100μg/mL)。保存产品时请避免反复冻融反复冻融，冰晶会破坏重组蛋白的空间结构，导致蛋白变性形成多聚体或使蛋白构象发生改变，从而降低蛋白的结合能力，也加快了蛋白的降解速度。MCEMasterofBioactiveMoleculesCONTENTS细胞因子和生长因子 2CytokinesandGrowthFactors免疫检查点蛋白4ImmuneCheckpointProteinsCAR-T相关蛋白6CAR-TRelatedProteinsCD抗原7CDAntigensFc受体蛋白8FcReceptorProteins受体蛋白9ReceptorProteins酶&调节子10Enzymes&Regulators补体系统相关蛋白11ComplementSystemRelatedProteins泛素相关蛋白12

病毒蛋白 13ViralProteins生物素标记蛋白 14BiotinylatedProteinsGMP级蛋白15GMP-gradeProteins无动物成分重组蛋白 15Animal-freeRecombinantProteins重组蛋白定制服务 16RecombinantProteinExpressionService客户验证 17CustomerValidation更多文献引用情况 18PublicationsCitingUseofMCERecombinantProteinsUbiquitinRelatedProteinswww.MedChemEInhibitors • ScreeningLibraries • Proteins细胞因子和生长因子细胞因子(Cytokines)是一类由多种免疫细胞(如巨噬细胞、淋巴细胞和肥大细胞等)以及某些非免疫细胞(如内皮细胞、表皮细胞等)分泌的低分子量可溶性蛋白/多肽/糖蛋白。细胞因子包括白介素、趋化因子、干扰素等，它们在调节细胞生长、分化和激活中发挥重要作用，并参与先天和适应性免疫反应[1][2][3]。生长因子(GrowthFactors)是一类可溶性信号分子，通常为分泌蛋白或甾体激素，能够在发育和组织愈合过程中刺激细胞增殖、迁移、分化和多细胞形态发生[4][5]。“生长因子”和“细胞因子”常通用。某些多肽生长因子为细胞因子，影响细胞生长分化信号途径的细胞因子被认为是生长因子，如GM-CSF[5][6]。MCE提供高纯度、高活性细胞因子，助力您的研究！HumanIGF-I(HY-P7018)HumanIGF-I(HY-P7018)ThepurityofhumanIGF-Iisgreaterthan95%asanalyzedbySDSunderreducing(R)conditions.kDa M 120806040302010HumanEGF(HY-P7109)

TheED50ofhumanIGF-Iis<5.0ng/mlasmeasuredbyFDC-P1cells,correspondingtoaspecificactivityof>2.0×10units/mg.0.6OD4900.40.2Net0.0101010-210-11010101010IGF-I,Human(ng/mL)HumanEGF(HY-P7109)ThepurityofhumanEGFisgreaterthan95%asanalyzedbySDSunderreducing(R)conditions.kDa M R120806040302010Cat.No.ProductNameHY-P7080IL-4HY-P7223IL-6RalphaHY-P7025IFN-gammaHY-P7287SDF-1beta/CXCL12

TheED50ofhumanEGFis<0.2ng/mLasmeasuredbymurineBALB/c3T3cells,correspondingtoaspecificactivityof5.0×10units/mg.0.6OD490nm0.40.2Net0.01010101010EGF,Human(ng/mL)SpeciesSourceTagMouseMammalianCellsTag-freeHumanMammalianCellsTag-freeHumanE.coliTag-freeHumanE.coliTag-free2MCEMasterofBioactiveMoleculesCat.No.ProductNameSpeciesSourceTagHY-P7058TNF-alpha/TNFSF2HumanE.coliTag-freeHY-P7085M-CSFMouseE.coliTag-freeHY-P7118TGFbeta1HumanMammalianCellsTag-freeHY-P7086NogginMouseMammalianCellsTag-freeHY-P7007BMP-4HumanE.coliTag-freeHY-P70593FibronectinHumanE.coliTag-freeHY-P70453Wnt3aHumanMammalianCellsTag-freeHY-P7109EGFHumanE.coliTag-freeHY-P70311ActivinAHuman/Mouse/RatMammalianCellsTag-freeHY-P7319AITRL/TNFSF18MouseE.coliTag-freeHY-P75509Angiopoietin-2CanineMammalianCellsN-HisHY-P72650FGF-21CynomolgusMammalianCellsC-HisHY-P72106BMP1HumanE.coliN-HisHY-P71827Adiponectin/ADIPOQBovineP.pastorisN-HisHY-P7008BMP-7HumanE.coliTag-freeHY-P7257CCL4HumanE.coliTag-freeHY-P70450CCL5HumanE.coliTag-freeHY-P7143CCL6MouseE.coliTag-freeHY-P7772CCL9MouseE.coliTag-freeHY-P70138DLK-1HumanMammalianCellsC-HisHY-P7004FGFbasic/bFGFHumanE.coliTag-freeHY-P7170FGF-10MouseE.coliTag-freeHY-P7346FGF-8HumanE.coliTag-freeHY-P7015AG-CSFHumanMammalianCellsTag-freeHY-P7016GM-CSFHumanMammalianCellsTag-freeHY-P7017HB-EGFHumanE.coliTag-freeHY-P7018IGF-IHumanE.coliTag-freeHY-P7368IGFBP-2HumanMammalianCellsC-HisHY-P7027IL-1alphaHumanE.coliTag-freeHY-P7030AIL-10HumanMammalianCellsTag-freeHY-P7049LIFHumanE.coliTag-freeHY-P7051ANogginHumanMammalianCellsTag-freeHY-P70781SCFHumanE.coliTag-freeHY-P70467SHHHumanE.coliTag-free3www.MedChemEInhibitors • ScreeningLibraries • Proteins免疫检查点蛋白免疫检查点(ImmuneCheckpoints)分子是对免疫应答发挥抑制或刺激作用的配体-受体对，它们在维持免疫稳态和免疫耐受中起着重要的调节作用。大多数被报道的免疫检查点蛋白在适应性免疫系统（特别是T细胞）和先天免疫系统的细胞上表达。一些肿瘤细胞通过结合共抑制受体分子，限制正常的抗肿瘤免疫反应，从而帮助免疫逃逸。癌症的免疫检查点治疗策略包括了靶向这些调控途径，以恢复免疫细胞的抗肿瘤功能。目前，研究最广泛的抑制性免疫检查点包括CTLA-4和PD-1/PD-L1等[7][8]。A2aRVISTAAdenosineB7-H3PDL1/PDL2A2aRGalectin-9??CD80/CD86PD1TIM-3CD80/CD86CTLA-4OX40LCD28T-CellCD40OX40CD40LB7RP1ICOSCD70CD27HVEMBTLAAPCorLAG-3TumorCellTCRMHCl/llKIRGITRGITRL4-1BBCD226/TIGIT4-1BBLCD155/CD112OtherpotentialpathwaysIL10 CD47IDO TLRTGF-β ArginaseInhibitoryPathwaysStimulatoryPathwaysFigure1.常见的免疫检查点刺激途径和抑制途径[9]4MCEMasterofBioactiveMoleculesCat.No.ProductNameSpeciesSourceTagHY-P70691CTLA-4HumanMammalianCellsC-GSTHY-P70632PD-L1MouseMammalianCellsC-HisHY-P7395PD-1HumanMammalianCellsC-hFcHY-P70482TIM3HumanMammalianCellsC-HisHY-P70722LAG-3HumanMammalianCellsC-HisHY-P70624TIGITHumanMammalianCellsC-HisHY-P7327CD276/B7-H3HumanMammalianCellsC-HisHY-P74464-1BBL/TNFSF9MouseMammalianCellsN-HisHY-P7144CD40L/CD154/TRAPHumanE.coliTag-freeHY-P70652CD276/B7-H3HumanMammalianCellsC-HisHY-P7678BTLA/CD272HumanMammalianCellsC-HisHY-P7685BTN3A3HumanMammalianCellsC-HisHY-P70535Galectin-9HumanMammalianCellsC-HisHY-P7366HVEMHumanSf9insectCellsC-hFcHY-P70494Nectin-1HumanMammalianCellsC-HisHY-P70807PVR/CD155HumanMammalianCellsC-HisHY-P73370PD-L2RatMammalianCellsC-hFcHY-P77879VISTAHumanMammalianCellsC-hFcHY-P75610CD137/4-1BBCanineMammalianCellsC-HisHY-P7394OX40/TNFRSF4HumanMammalianCellsC-HisHY-P77457OX40Ligand/TNFSF4CynomolgusMammalianCellsN-mFcHY-P73499CD40HumanMammalianCellsC-HisHY-P73306Nectin-3HumanMammalianCellsC-HisHY-P71248PVRIGHumanMammalianCellsC-mFcHY-P76070SIRPalphaMouseMammalianCellsC-HisHY-P73121IDOHumanE.coliTag-freeHY-P76396ICOSHumanMammalianCellsC-His-hFcHY-P77575ICOSLGCynomolgusMammalianCellsC-HisHY-P72353CD28Human/CynomolgusMammalianCellsC-Fc-AviHY-P72033LIGHTHumanMammalianCellsN-hFc-MycHY-P73076GITRHumanMammalianCellsC-HisHY-P7318GITRL/AITRLHumanE.coliTag-freeHY-P72887CD200HumanMammalianCellsC-hFcHY-P76780CD200R1CynomolgusMammalianCellsC-His5www.MedChemEInhibitors • ScreeningLibraries • ProteinsCAR-T相关蛋白嵌合抗原受体T(CAR-T)细胞是通过基因工程表达嵌合抗原受体的T细胞，能够特异性地靶向抗原并杀死肿瘤细胞[10][11]。CAR-T细胞能够通过单链可变区片段(scFv)结合域特异性识别靶抗原，以不依赖于主要组织相容性复合体(MHC)的方式激活，从而靶向杀伤肿瘤细胞，因此靶标抗原的选择对CAR-T细胞的特异性杀伤作用至关重要[12]。靶点蛋白结合流式细胞法是常用的CAR-T转染阳性率检测方案。目前MCE重组蛋白已覆盖BCMA、EGFR、HER2、CD22、CD19、CD138、ROR1、CD123、Mesothelin等40+个CAR-T热门靶标。TumorCellScFvHingeRegionCostimulatoryDomainCD3ZetaFigure2.嵌合抗原受体示意图[13]Cat.No.ProductNameSpeciesSourceTagHY-P7656BCMA/TNFRSF17MouseMammalianCellsC-FcHY-P72019Siglec-2/CD22HumanMammalianCellsC-HisHY-P70505CD19HumanMammalianCellsC-FcHY-P70731CD38HumanMammalianCellsC-HisHY-P70148MesothelinHumanMammalianCellsC-HisHY-P70189EGFRvIII`HumanMammalianCellsC-HisHY-P70125CD276/B7-H3CynomolgusMammalianCellsC-HisHY-P70155EpCAM/TROP1HumanMammalianCellsC-FcHY-P70759HER2/CD340HumanMammalianCellsC-FcHY-P70301Mucin-1HumanMammalianCellsC-FcHY-P71031Siglec-6HumanMammalianCellsC-FcHY-P70487Glypican-3/GPC3HumanMammalianCellsC-HisHY-P70296FolatereceptoralphaHumanMammalianCellsC-HisHY-P70497CD7HumanMammalianCellsC-His6MCEMasterofBioactiveMoleculesCD抗原分化簇抗原(CDAntigens)指通常用于白细胞表面分子的鉴定和研究的分化抗原群/分化抗原簇。一些CD蛋白通常充当细胞间/细胞基质粘附分子、细胞因子受体、离子通道或营养转运蛋白，它们在免疫系统功能中发挥多种作用。利用CD抗原作为细胞标记物，通过荧光标记抗体和流式细胞术结合的方法，对特定的白细胞群和淋巴细胞亚群进行鉴定和分选[14][15]。CD45R CD45RCD28 CD28CD2 CD2CD3THTCCD3TCRTCRCD4CD8LFA1LFA1Figure3.辅助T细胞和毒性T细胞表面抗原标志物[16]Cat.No.ProductNameSpeciesSourceTagHY-P70090CD3epsilonCynomolgusMammalianCellsC-FcHY-P72702CD8betaHumanMammalianCellsN-HisHY-P70486CD28Human/CynomolgusMammalianCellsN-HisHY-P7321B7-1/CD80HumanMammalianCellsC-hFcHY-P70029Basigin/CD147HumanMammalianCellsC-HisHY-P70507CD44HumanMammalianCellsC-HisHY-P7679BTLA/CD272HumanMammalianCellsC-FcHY-P7780Nectin-2/CD112HumanMammalianCellsC-HisHY-P7785CD127/IL-7RAHumanMammalianCellsC-Fc-HisHY-P70550CD137/4-1BBCynomolgusMammalianCellsC-HisHY-P7799CD160MouseMammalianCellsC-HisHY-P70479CD40HumanMammalianCellsC-HisHY-P7819CD207HumanMammalianCellsN-His7www.MedChemEInhibitors • ScreeningLibraries • ProteinsFc受体蛋白免疫球蛋白Fc受体(FcReceptors)是表达于大多数先天性和适应性免疫细胞的膜蛋白，属于免疫球蛋白超家族。Fc与抗体(IgA、IgD、IgE、IgG和IgM)的FabFc区域相互作用，发挥多种生物效应，例如通过释放细胞因子或吞噬作用调节免疫反应。这些Fc受体包括：IgG(FcγRI/CD64、FcγRII/CD32、FcγRIII/CD16、FcRn)、IgE(FcεRI、FcεRII)、IgA(FcαRI/CD89)、IgM(FcμR)、IgD(FcδR)和Fcα/μR(IgA/IgM)等[17][18]。AntibodyD2FcReceptorEffectorCellFigure4.Fc受体与抗体包被的微生物病原体相互作用[19]

CDRVariableRegionVHVLCH1Fab-FragmentCLLightChain(korλ)CH2Fc-FragmentCH3HeavyChainsFigure5.IgG结构图[20]Cat.No.ProductNameSpeciesSourceTagHY-P70708FcgammaRIII/CD16MouseMammalianCellsC-HisHY-P70490FcgammaRIIIA/CD16aHumanMammalianCellsC-HisHY-P72657FCAR/CD89HumanMammalianCellsC-HisHY-P70711FcgammaRIIA/CD32aRatMammalianCellsC-HisHY-P70669CD64HumanMammalianCellsC-HisHY-P70601FCRNHumanMammalianCellsC-HisHY-P72748CD23/FcepsilonRIIHumanMammalianCellsN-HisHY-P76800FcgammaRIIB/CD32bCynomolgusMammalianCellsC-HisHY-P75204FcgammaRIIIB/CD16bHumanE.coliBiotinylatedHY-P72191FcepsilonRIA/FCER1AHumanE.coliHis-SUMOHY-P77363FCAMR/CD351MouseMammalianCellsC-HisHY-P70251IgG3FcMouseMammalianCellsTag-freeHY-P72603IgG2AFcMouseMammalianCellsTag-free8MCEMasterofBioactiveMolecules受体蛋白受体(Receptors)是位于细胞表面或胞内的蛋白，它们与膜上或胞内的多种信号传导系统关联，通过特异性地识别和结合配体调节细胞对细胞/组织微环境的反应[21]。细胞受体种类繁多，大体分为内部受体和细胞表面受体。细胞表面受体包括三大类：离子通道受体、G-蛋白偶联受体(GPCRs)和酶联受体[22]。目前，受体已经成为一大类药物靶点，例如GPCRs、细胞因子受体[23]。GLP-1-BasedDrugsGLP-1RTNF-αPalmitateLPSPKCPMAACIFN-γNLRP3cAMP/CaOx-LDLERK1/2CAMKKβPKACaspase-1pAMPK

pro-IL-1βIL-1βNF-B STAT1 NucleusCytokineProductionTNF-α,IL-6,IFN-γ,IL-1β,IL-17,IL-2

AdhesionMoleculesVCAM-1,ICAM-1,E-selectinAnti-inflammation

ChemokinesCXCL10,MCP-1InflammationInducerFigure6.基于GLP1-GLP1R分子信号的药物抗炎作用机制[24]Cat.No.ProductNameSpeciesSourceTagHY-P70552VEGFR-2HumanMammalianCellsC-HisHY-P70714HGFRHumanMammalianCellsC-HisHY-P70352GLP1RHumanMammalianCellsC-FcHY-P70793TrkBHumanMammalianCellsC-HisHY-P7308TrkAHumanMammalianCellsTag-freeHY-P70179LIR-1/LILRB1HumanMammalianCellsC-HisHY-P7485ActivinRIB/ALK-4HumanMammalianCellsC-HisHY-P7467AGERHumanMammalianCellsC-HisHY-P71580GFRALHumanE.coliN-His-SUMOHY-P72198FSHRHumanE.coliN-HisHY-P70057TYRO3/DTKMouseMammalianCellsC-His9www.MedChemEInhibitors • ScreeningLibraries • Proteins酶&调节子酶类(Enzymes)是在生物系统内发现的生物催化剂，能催化特定的生化过程。大多数酶是具有催化活性的蛋白质，部分RNA分子(核酶)也具有催化活性。发生在生物机体内的生物过程大多数是由酶调节的。许多遗传性的人类疾病，如白化病和苯丙酮尿症，都是由于缺乏特殊的酶引起的。酶催化反应能被内源或外源抑制剂抑制。许多毒素或活性药物通过抑制特定的酶促反应发挥作用[25][26][27]。1EnzymeAvailableSubstrate(sucrose)withEmptyActiveSiteActiveSite2SubstrateBindstoEnzymewithInducedFitGlucoseEnzyme(sucrase)FructoseH2O4Productsare3SubstrateisConvertedReleasedtoProductsFigure7.酶的催化作用[28]Cat.No.ProductNameSpeciesSourceTagHY-P7745CathepsinAHumanMammalianCellsC-HisHY-P7442ACE2HumanMammalianCellsC-FcHY-P70351MMP-9MouseMammalianCellsC-HisHY-P70051AminopeptidaseN/CD13MouseMammalianCellsC-HisHY-P7474ALDH1A1HumanE.coliN-HisHY-P7734CarboxypeptidaseB1/CPB1HumanMammalianCellsC-HisHY-P70005CTRB1HumanMammalianCellsC-HisHY-P70010CD73/5'-NucleotidaseHumanMammalianCellsC-HisHY-P7452ACOT13HumanMammalianCellsC-HisHY-P70221Acyl-proteinthioesterase2/LYPLA2HumanE.coliC-HisHY-P7479Aldose1-epimerase/GALMHumanE.coliC-HisHY-P72076ALOX12HumanE.coliN-HisHY-P70260Alpha-enolase/Enolase1HumanE.coliC-HisHY-P73090GSK-3betaMouseE.coliN-HisHY-P7503AngiogeninHumanE.coliTag-freeHY-P73267Kininogen-1MouseMammalianCellsC-HisHY-P7602Arginase-1HumanMammalianCellsC-His10MCEMasterofBioactiveMolecules补体系统相关蛋白补体系统(ComplementSystem)是先天免疫的主要成分之一，由可溶性血浆蛋白、膜型补体调节蛋白和补体受体组成，存在于血浆、组织、细胞表面，甚至细胞内。补体系统主要有三条激活途径：经典途径(CP)、凝集素途径(LP)、旁路途径(AP)。这些途径依赖不同的起始分子激活，但最后产生相同的终端效应分子：过敏性毒素(C4a/C3a/C5a)、膜攻击复合物(MAC)和调理素(如C3b)。补体系统在防御病原体和维持宿主稳态中起着关键作用[29]。ComplementSystemClassicalPathwayLectinPathwayC1qMBLAg-AbComplexesLectinBindsMannoseonPathogensC1,MASPsC4,C4C2C2CR1C3ConvertaseFcRC3C3bOpsonizationC5ConvertasePhagocytosisC5C5bC6,C7,C8C9C5b-9

AlternativePathwayPathogens,InjuredTissueC3bC3bC3aRC3aC5aRInflammationC3aRC5aC5aRMACCellLysisandActivationFigure8.三种补体系统激活途径[30]Cat.No.ProductNameSpeciesSourceTagHY-P7863ComplementC3/C3aMouseE.coliTag-freeHY-P7864ComplementC5/C5aHumanE.coliTag-freeHY-P70031CFHR2HumanMammalianCellsC-HisHY-P70055CFHR1HumanMammalianCellsC-HisHY-P70102CFHR5HumanMammalianCellsC-HisHY-P7692HABP1/C1QBPHumanE.coliC-HisHY-P71718C1QAMouseP.pastorisN-HisHY-P71423VSIG4HumanMammalianCellsC-FcHY-P75395CD55/DAFHumanMammalianCellsC-HisHY-P7890ComplementfactorH/CFHHumanMammalianCellsC-HisHY-P74371C7/ComplementcomponentC7HumanMammalianCellsC-HisHY-P74375C2/ComplementC2HumanMammalianCellsC-HisHY-P74614ProteinS/PROS1HumanMammalianCellsC-HisHY-P75463C1s-AsubcomponentMouseMammalianCellsC-HisHY-P75464C1QBHumanSf9InsectCellsC-His11www.MedChemEInhibitors • ScreeningLibraries • Proteins泛素相关蛋白泛素(Ubiquitin)是真核细胞中广泛表达的一种高度保守的小分子蛋白，由76个氨基酸组成。泛素共价结合到靶蛋白上被称为“泛素化”，泛素化是一种重要的蛋白质翻译后修饰(PTM)，在控制蛋白的稳定和功能中起重要作用，并调节了多种细胞过程，例如细胞分裂和分化、环境应激反应、免疫反应、DNA修复和凋亡。著名的泛素-蛋白酶体途径(UPP)是重要的胞质蛋白降解途径，能选择性降解各种形式的受损蛋白。泛素化修饰由三类酶催化完成：1)泛素激活酶E1激活泛素蛋白；2)激活的泛素分子被转移到泛素结合酶E2的Cys残基上；3)泛素由E3连接酶连接到靶蛋白的Lys残基上[31][32][33]。ATPPP1UbE1UbAMPSubstrateCysDUBAMPLysUbUbUbE1SubstrateUbCysUbE3E2E2E2E3SubstrateCysE3UbLysE1SubstrateFigure9.蛋白泛素化过程[34]Cat.No.ProductNameSpeciesSourceTagHY-P70843NEDD8HumanE.coliTag-freeHY-P71016UBE2CHumanE.coliN-HisHY-P71020UBE2L6HumanE.coliC-HisHY-P71096UCH-L1HumanE.coliC-HisHY-P71101UBBHumanE.coliTag-freeHY-P70974SUMO2HumanE.coliN-HisHY-P7617ATG3HumanE.coliTag-freeHY-P71182OTUB2HumanE.coliN-GSTHY-P71396UBAP1HumanE.coliC-HisHY-P73537XIAPHumanE.coliN-HisHY-P71643SAE1HumanE.coliN-GSTHY-P70149ISG15/UCRPHumanE.coliC-HisHY-P71395UBA5HumanE.coliN-HisHY-P71402UBE2HHumanE.coliN-GSTHY-P71407UBE2R2HumanE.coliN-His12MCEMasterofBioactiveMolecules病毒蛋白一个完整的病毒粒子由其遗传物质(DNA或RNA)和蛋白外壳构成。构成成熟的病毒粒子的蛋白，即病毒结构蛋白，可能包括核衣壳核心蛋白(Gag蛋白)、病毒颗粒内包装的酶(Pol蛋白)和膜组分(Env蛋白)。病毒蛋白还包括非结构蛋白、调节蛋白和辅助蛋白，在感染细胞时，这些蛋白在病毒复制以及组装调节中发挥了重要作用[35][36][37]。MCE提供700+种包括了流感病毒、新型冠状病毒(SARS-CoV-2)、人类免疫缺陷病毒(HIV)、肝炎病毒等病毒相关的重组蛋白，助力病毒研究。NucleocapsidProtein(N)MembraneGlycoprotein(M)SpikeProtein(S)EnvelopeProtein(E)RNAFigure10.SARS-CoV-2简要结构图[38]Cat.No.ProductNameSpeciesSourceTagHY-P74293C-likeProteinaseSARS-CoV-2E.coliN-HisHY-P7437NucleocapsidSARS-CoV-2E.coliN-HisHY-P7436S1ProteinSARS-CoV-2MammalianCellsTag-freeHY-P70127NSP1SARS-CoV-2E.coliC-HisHY-P73290Spike/S1MERS-CoVSf9insectCellsC-HisHY-P70907Envelopeglycoproteingp120HIV-1MammalianCellsC-HisHY-P74883gp140HIV-1MammalianCellsC-FcHY-P70015B18RVacciniavirusMammalianCellsC-HisHY-P71478FusionglycoproteinF0/FHRSVAE.coliN-His,B2MHY-P73232HA/HemagglutininInfluenzavirusMammalianCellsC-HisHY-P73239NA/NeuraminidaseInfluenzavirusMammalianCellsTag-freeHY-P73533MembraneproteinZikavirusMammalianCellsC-FcHY-P73738NS1ProteinDenguevirusMammalianCellsN-HisHY-P74188E/EnvelopeProteinWestNileVirusP.pastorisC-HisHY-P74354CapsidproteinHepatitisvirusE.coliC-His13www.MedChemEInhibitors • ScreeningLibraries • Proteins生物素标记蛋白生物素化(Biotinylation)是将生物素以共价键连接到分子(如氨基酸或蛋白质)上的过程。由于生物素很小，生物素化通常快速且具有特异性，不易干扰分子固有功能。生物素与链霉亲和素或抗生物素蛋白的结合具有极高的亲和力、快速性和高特异性，因此，生物素化蛋白作为一种分子工具广泛用于生物医学领域。AviTag™是常用的融合标签，Avi-Tag是一种由15个氨基酸残基组成的小标签，能被大肠杆菌生物素连接酶BirA有效生物素化，可添加在蛋白的N或C端，实现蛋白的生物素化，常用于蛋白纯化或显影等[39]。AviTagAviTagProtein—GLNDIFEAQKIEWHEProtein—GLNDIFEAQKIEWHEN3H+BirANH+OOOATPAMP+PPiSHHNHSHHNONHHHNBiotinOBiotinFigure11.Avi标记的靶蛋白被BirA生物素化[40]Cat.No.ProductNameSpeciesSourceTagHY-P70546PCSK9HumanMammalianCellsC-His-HA-AviHY-P70721PD-L1HumanMammalianCellsC-Fc-AviHY-P70767Siglec-15HumanMammalianCellsC-Fc-AviHY-P70768CD79BHumanMammalianCellsC-His-AviHY-P70769ACE2HumanMammalianCellsC-His-AviHY-P70770TROP-2HumanMammalianCellsC-His-AviHY-P71175NTNG1HumanMammalianCellsC-Avi-HisHY-P71056TGFbeta1HumanMammalianCellsN-AviHY-P71421VEGFR-2HumanMammalianCellsC-His-AviHY-P73309Neuropilin-1HumanMammalianCellsC-His-AviHY-P73485XIAPHumanE.coliN-AviHY-P728824-1BBR/TNFRSF9HumanMammalianCellsC-hFc-AviHY-P72909CD40HumanMammalianCellsC-hFc-AviHY-P72885FcgammaRIIIA/CD16aHumanMammalianCellsC-His-AviHY-P72343BTN1A1HumanMammalianCellsC-His-AviHY-P72389LAG-3HumanMammalianCellsC-His-Avi14MCEMasterofBioactiveMoleculesGMP级蛋白GMP(生产质量管理规范)重组蛋白在特定的指导下生产，以确保产品质量和一致性。MCE提供的GMP级别重组蛋白是在独立的QA监督下生产，并具有质量记录和完全可追溯性。Cat.No.ProductNameSpeciesSourceTagHY-P70637GGMPTPOHumanMammaliancellsC-HisHY-P70757GGMPSCFHumanMammaliancellsC-HisHY-P70454GGMPIL-1alphaHumanE.coliTag-freeHY-P7044GGMPIL-6HumanE.coliTag-freeHY-P7032GGMPIL-12HumanMammalianCellsTag-freeHY-P70760GGMPIL-18HumanMammalianCellsC-HisHY-P7038GGMPIL-21HumanE.coliTag-freeHY-P78551GMPIL-2HumanE.coliTag-freeHY-P7055GGMPPDGF-BBHumanE.coliTag-free无动物成分重组蛋白无动物成分(Animal-free)重组蛋白，即重组蛋白生产过程中不使用任何动物来源的原料和试剂，产品接触的所有材料均不含任何动物成分。在无动物成分条件下生产的与使用标准实验室技术生产的蛋白具有相同的生物活性，同时避免痕量的动物成分、动物病原体和抗原对实验造成影响。Cat.No.ProductNameSpeciesSourceTagHY-P700017AFAnimal-FreeBAFFHumanE.coliC-HisHY-P700018AFAnimal-Freebeta-NGFHumanE.coliC-HisHY-P700020AFAnimal-FreeBMP-11HumanE.coliC-HisHY-P700034AFAnimal-FreeCCL2HumanE.coliN-HisHY-P700037AFAnimal-FreeCD326HumanE.coliC-HisHY-P700042AFAnimal-FreeCXCL11HumanE.coliN-HisHY-P700043AFAnimal-FreeCXCL12HumanE.coliC-HisHY-P700045AFAnimal-FreeCXCL2HumanE.coliN-HisHY-P700056AFAnimal-FreeFGF-12HumanE.coliC-HisHY-P700066AFAnimal-FreeFGF-4HumanE.coliC-HisHY-P700083AFAnimal-FreeG-CSFHumanE.coliN-His15www.MedChemEInhibitors • ScreeningLibraries • Proteins重组蛋白定制服务从基因设计到蛋白制备的一站式服务自主研发多种高表达载体与菌株专业的重组蛋白技术研发团队提供毫克至克级高纯度蛋白定制服务提供蛋白在生物研究应用中专业的技术支持

基因合成密码子优化交付蛋白

表达载体构建菌株筛选蛋白表达服务内容

蛋白纯化MCE提供从基因到蛋白的一站式服务，客户可以根据自身需求灵活性选择服务或个性化定制服务。服务内容包括：服务项目服务内容服务周期交付内容基因合成及亚克隆2-3周构建报告大肠杆菌表达系统表达检测1周表达检测报告放大表达及蛋白纯化1-3周纯化蛋白/纯化报告基因合成及亚克隆2-3周构建报告酵母表达株电转1-2周阳性菌株酵母细胞表达系统高拷贝细胞株筛选（可选）2周高拷贝菌株酵母阳性细胞株表达检测3周表达检测报告放大表达及蛋白纯化3-5周目的蛋白/纯化报告基因合成及亚克隆2-3周构建报告哺乳动物细胞表达系统瞬时转染及表达小试2-3周蛋白表达报告放大培养及蛋白纯化3-5周目的蛋白/纯化报告基因合成及亚克隆2-3周构建报告制备Bacmid1-2周制备报告昆虫细胞表达系统杆粒及P2代病毒制备2-3周表达鉴定报告转染及表达小试1-2周蛋白表达检测报告放大培养及蛋白纯化3-4周目的蛋白/纯化报告16MCEMasterofBioactiveMolecules客户验证诱导小鼠肺成纤维细胞纤维化TGF-β1(HY-P7117)刺激原代小鼠肺成纤维细胞，通过qRT-PCR和WB检测PTPN13和NOX4表达水平，确定其与细胞纤维化密切相关[41]。ControlTGF-β1PTPN13NOX4expression10**expression8**86mRNA6mRNA44Relative2Relative200

ControlTGF-β1ControlTGF-β1CollagenI139kDaPTPN13310kDaα-SMA42kDaNOX455kDaGAPDH37kDaGAPDH37kDa构建肾脏类器官从小鼠肾脏组织中提取细胞，利用微流体技术结合3D打印机制作类器官珠，辅以Noggin(HY-P7086)、R-spondin-1(HY-P7114)、FGF-4(HY-P7014)、FGF-basic(HY-P7066)、SB-431542(HY-10431)、Laduviglusib(HY-10182)培养，7天后，得到数千个大小、形状和成分具有高度重复性的肾脏类器官[42]。OrganoidGeneratingKidneyOrganoidBeadsKidneyOrganoidsMatrigel+CellsDay1Day7QDs/Drugs7days24hrsOrganoidCulturingCulturingHighThroughputScreeningOil4°CKidneyOrganoidBeads200μm200μmCultureDish37°CIFN-γ诱导蛋白16(IFI16)的积累通过Westernblotting检测IFN-γ(HY-P7025)处理人肺腺癌上皮(A549)细胞18小时后，蛋白16(IFI16)的积累，通过免疫染色检测IFN-γ诱导16小时后IFI16的细胞定位[43]。HumanIFN-γ(ng/mL)0505001000IFI16—100(kDa)—70β-actin—40

NegativeControl IFN-γ(500ng/mL)DAPI/IFI1617MCEMasterofBioactiveMoleculesPublicationsCitingUseofMCEProductsNature.2023Jun;618(7964):374-382.Nature.2023Jun;618(7966):862-870.Nature.2023Jun;618(7965):590-597.Nature.2023Jun;618(7963):180-187.Nature.2023May;617(7962):818-826Science.2023Jun9;380(6649):eabo2296.Science.2022Dec2;378(6623):eabo5503.Science.2022Nov18;378(6621):eabq7361.Science.2022Oct14;378(6616):eabq0132.Science.2022Jul8;377(6602):eabg9302Cell.2023Jun22;186(13):2823-2838.e20.Cell.2023May11;186(10):2144-2159.e22.Cell.2023Mar30;186(7):1352-1368.e18.Cell.2023Mar2;186(5):1026-1038.e20.Cell.2023Feb16;186(4):850-863.e16MCEGlobalPartnerswww.MedChemEInhibitors • ScreeningLibraries • 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