Recombinant Human CDH17/Cadherin 17 (His Tag)

來源: 發(fā)布時(shí)間:2024-06-04

Endo H糖苷內(nèi)切酶H:結(jié)構(gòu)、功能及其在糖生物學(xué)中的應(yīng)用摘要Endo H糖苷內(nèi)切酶H(Endo H)是一種專門切割N-連接糖鏈的內(nèi)切酶,用于糖生物學(xué)和蛋白質(zhì)工程領(lǐng)域。本文將探討Endo H的結(jié)構(gòu)特性、催化機(jī)制、以及其在研究和臨床應(yīng)用中的潛力。引言N-連接糖基化是一種在真核細(xì)胞中普遍存在的蛋白質(zhì)修飾方式,涉及將糖鏈添加到蛋白質(zhì)的天冬酰胺殘基上。Endo H作為一種內(nèi)切酶,能夠特異性地識(shí)別并切割N-連接糖鏈中的β-N-乙酰葡糖胺苷鍵,從而在蛋白質(zhì)工程和生物制藥中發(fā)揮重要作用。Endo H的結(jié)構(gòu)特性Endo H通常來源于流感嗜血桿菌(Hemophilus influenzae),其分子質(zhì)量約為50 kDa。該酶由兩個(gè)結(jié)構(gòu)域組成:一個(gè)負(fù)責(zé)識(shí)別糖鏈的催化結(jié)構(gòu)域和一個(gè)有助于酶穩(wěn)定性的非催化結(jié)構(gòu)域。催化機(jī)制Endo H的催化機(jī)制涉及對(duì)N-連接糖鏈的特異性識(shí)別和切割。酶的活性位點(diǎn)含有多個(gè)氨基酸殘基,這些殘基與糖鏈的特定結(jié)構(gòu)相互作用,導(dǎo)致酶對(duì)底物的高特異性。Endo H催化的糖苷鍵斷裂是一水解反應(yīng),需要水分子的參與。CB1受體分布于大腦和外周神經(jīng)系統(tǒng)中,與多種生理功能有關(guān),包括疼痛感知、食欲調(diào)節(jié)、情緒調(diào)節(jié)。Recombinant Human CDH17/Cadherin 17 (His Tag)

Recombinant Human CDH17/Cadherin 17 (His Tag),標(biāo)準(zhǔn)物質(zhì)

SARS-CoV-2,whichcausestheglobalpandemiccoronavirusdisease2019(Covid-19),belongstoafamilyofvirusesknownascoronavirusesthatalsoincludeMERS?CoVandSARS-CoV-1.Coronavirusesarecommonlycomprisedoffourstructuralproteins:Spikeprotein(S),Envelopeprotein(E),Membraneprotein(M)andNucleocapsidprotein(N).TheSARS-CoV-2Sproteinisaglycoproteinthatmediatesmembranefusionandviralentry.TheSproteinishomotrimeric,witheach~180-kDamonomerconsistingoftwosubunits,S1andS2.TheRBDofSARS-CoV-2bindsametallopeptidase,angiotensin-convertingenzyme2(ACE-2).BeforebindingtotheACE-2receptor,structuralanalysisoftheS1trimershowsthatonlyoneofthethreeRBDdomainsisinthe"up"conformation.Thisisanunstableandtransientstatethatpassesbetweentrimericsubunitsbutisneverthelessanexposedstatetobetargetedforneutralizingantibodytherapy.PolyclonalantibodiestotheRBDoftheSARS-CoV-2proteinhavebeenshowntoinhibitinteractionwiththeACE-2receptor,confirmingRBDasanattractivetargetforvaccinationsorantiviraltherapy.Influenza HA (518-526)隨著醫(yī)藥科技的發(fā)展,透明質(zhì)酸在醫(yī)藥這方面的應(yīng)用將越來越多,包括作為藥物載體和組織工程的基質(zhì)。

Recombinant Human CDH17/Cadherin 17 (His Tag),標(biāo)準(zhǔn)物質(zhì)

RecombinantBiotinylatedHumanHBV(HLA-A*02:01)Protein,His-AviTag性能參數(shù)表達(dá)區(qū)間及表達(dá)系統(tǒng)(Source)RecombinantBiotinylatedHumanHBV(HLA-A*02:01)ProteinisexpressedfromHEK293withHistagandAvitagattheC-terminalItcontainsGly25-Thr305(HLA-A*02:01),Ile21-Met119(B2M)andFLLTRILTIpeptide.[Accession|A0A140T913(HLA-A*02:01)&P61769(B2M)&FLLTRILTIpeptide]分子量大?。∕olecularWeight)TheproteinhasapredictedMWof50.5kDa.Duetoglycosylation,theproteinmigratesto52-62kDabasedonTris-BisPAGEresult.(Endotoxin)Lessthan1EUperμgbytheLALmethod.純度(Purity)>95%asdeterminedby>95%asdeterminedbyHPLC制劑(Formulation)Lyophilizedfrom0.22μmfilteredsolutioninPBS(pH7.4).Normally8%trehaloseisaddedasprotectantbeforelyophilization.重構(gòu)方法(Reconstitution)Centrifugetubesbeforeopening.Reconstitutingtoaconcentrationmorethan100μg/mlisrecommended.Dissolvethelyophilizedproteinindistilledwater.

Trop-2,alsoknownasepithelialglycoprotein-1antigen(EGP-1),isaproteinthatinhumansisencodedbytheTACSTD2gene.Mutationsofthisgeneresultingelatinousdrop-likecornealdystrophy,anautosomalrecessivedisordercharacterizedbyseverecornealamyloidosisleadingtoblindness.產(chǎn)品性質(zhì)別名EGP1;EGP-1;TROP2;GA733-1;gp50;T16;TACSTD2;TROP-2;M1S1;TACD2UniprotNo.P09758表達(dá)區(qū)間及表達(dá)系統(tǒng)RecombinantBiotinylatedHumanTROP-2/TACSTD2ProteinisexpressedfromHEK293cellswithHistagandAvitagattheC-terminal.ItcontainsHis27-Thr274.分子量TheproteinhasapredictedMWof30.5kDa.Duetoglycosylation,theproteinmigratesto46-55kDabasedonTris-BisPAGEresult.純度>95%asdeterminedbySDS-PAGEandHPLC活性ELISAData:ImmobilizedAnti-TROP-2Antibody,hFcTagat0.5μg/mL(100μL/well)ontheplate.DoseresponsecurveforBiotinylatedHumanTROP-2,HisTagwiththeEC50of24.1ng/mLdeterminedbyELISA.0EUper1μgoftheproteinbytheLALmethod.制劑Lyophilizedfrom0.22μmfilteredsolutioninPBS(pH7.4).Normally5%trehaloseisaddedasprotectantbeforelyophilization.全長(zhǎng)A2aR蛋白具有天然完整構(gòu)象,VLP(病毒樣顆粒)固有特征可以帶來更高的免疫原性。

Recombinant Human CDH17/Cadherin 17 (His Tag),標(biāo)準(zhǔn)物質(zhì)

IdeSProtease全稱免疫球蛋白G降解酶,酶活(Enzymeactivity)40U/μL酶活定義(UnitDefinition)在37℃條件下,反應(yīng)30min,剪切>95%的1μg重組單克隆IgG所需要的酶量定義為一個(gè)活性單位。儲(chǔ)存條件-25~-15℃保存,有效期1年。使用方法1.在消化液中加入適量IgG(加至5mg);2.在IgG樣本中加入IdeS蛋白酶(每1μgIgG加入1個(gè)單位的IdeS);3.將樣品置于37℃孵育30-60min。IdeS蛋白酶在中性pH或接近中性pH的緩沖中活性強(qiáng)。推薦反應(yīng)緩沖是50mM磷酸鈉,150mMNaCl(pH6.6),多數(shù)常見的生物緩沖也適用,比如Tris或PBS;注意:不在此pH范圍(例如乙酸鹽緩沖液)的緩沖也可能適用,但是孵育時(shí)間或酶量需要根據(jù)實(shí)際情況進(jìn)行優(yōu)化。注意事項(xiàng)1.IdeS不能識(shí)別切割小鼠IgG1/IgG2b,大鼠、豬、牛和山羊IgG,對(duì)小鼠IgG2a和IgG3具有中等酶切活性,酶切小鼠IgG2a和IgG3建議增加IdeS的用量(推薦用量為正常用量的5-10倍)。2.IdeS不能切割非IgG亞型的單抗分子,包括IgA、IgM、IgD和IgE。全長(zhǎng)膜蛋白由于其天然構(gòu)象,是跨膜蛋白藥物開發(fā)中的好的抗原。在細(xì)胞中的表達(dá)水平通常較低。Recombinant Human S100A14Protein,His Tag

跨膜蛋白在宿主細(xì)胞中的表達(dá)水平通常有點(diǎn)低,研發(fā)困難,對(duì)蛋白表達(dá)生產(chǎn)平臺(tái)技術(shù)要求極高。Recombinant Human CDH17/Cadherin 17 (His Tag)

RecombinantBiotinylatedHumanENPP-3Protein,His-AviTag性能參數(shù)分子別名(Synonyms)NPP3;E-NPP3;PD-Ibeta表達(dá)區(qū)間及表達(dá)系統(tǒng)(Source)BiotinylatedHumanENPP-3ProteinisexpressedfromHEK293withHistagandAvitagattheN-Terminus.ItcontainsLeu48-Ile875.[Accession|O14638]分子量大?。∕olecularWeight)TheproteinhasapredictedMWof99.97kDa.Duetoglycosylation,theproteinmigratesto110-130kDabasedonSDS-PAGEresult.(Endotoxin)Lessthan1EUperμgbytheLALmethod.純度(Purity)>95%asdeterminedbySDS-PAGEandHPLC.制劑(Formulation)Lyophilizedfrom0.22μmfilteredsolutioninPBS(pH7.4).Normally8%trehaloseisaddedasprotectantbeforelyophilization.重構(gòu)方法(Reconstitution)Centrifugethetubebeforeopening.Reconstitutingtoaconcentrationmorethan100μg/mlisrecommended.Dissolvethelyophilizedproteinindistilledwater.儲(chǔ)存條件Theproductshouldbestoredat-25~-15℃for1yearfromdateofreceipt.2-7days,2~8°Cundersterileconditionsafterreconstitution.3-6months,-85~-65℃understerileconditionsafterreconstitution.Recombinant Human CDH17/Cadherin 17 (His Tag)