当前位置 ： NEB >>> NEB/Protein Deglycosylation Mix II/P6044S/20 reactions
NEB/Protein Deglycosylation Mix II/P6044S/20 reactions
  • NEB/Protein Deglycosylation Mix II/P6044S/20 reactions

NEB/Protein Deglycosylation Mix II/P6044S/20 reactions

价格: ¥5880.00 市场价: 9800.00
打印产品信息
货号: P6044S-20reactions
品牌: NEB
规格
数量
库存(0)
特别 提示
代购产品：无质量问题不接受退换货，下单前请仔细核对信息。
下单后请及时联系客服核对商品价格，订单生效后再付款。
立即购买 加入购物车

收藏商品
资深产品顾问
咨询顾问

全国免费服务热线

4000-520-616

  • 自营商城 一站式服务
  • 厂家直采 剔除溢价
  • 品质甄选 正品保证
  • 严控流程 只做188精品
  • 极速物流 如约送货
  • 详情
  • 使用说明
  • 常见问题

    • Description:

      Glycosylationisoneofthemostcommonpost-translationalmodificationsofproteins,asshowninFigure1.N-linkedglycosylationoccurswhenglycansareattachedtoasparagineresiduesonthecoreprotein.O-linkedglycosylationoccurswhenglycansareattachedtoserineorthreonineresidues.Bothchemicalandenzymaticmethodsexistforremovingoligosaccharidesfromglycoproteins.However,chemicalmethodssuchasβ-eliminationwithmildalkaliormildhydrazinolysiscanbeharshandmayresultinincompletesugarremovalanddegradationoftheprotein;whereas,enzymaticmethodsaremuchgentlerandcanprovidecompletesugarremovalwithnoproteindegradation.

      PNGaseFisthemosteffectiveenzymaticmethodforremovingalmostallN-linkedoligosaccharidesfromglycoproteins.PNGaseFdigestiondeaminatestheaspargineresiduetoasparticacid,andleavestheoligosaccharideintact,keepingitsuitableforfurtheranalysis.Oligosaccharidescontainingafucoseα(1-3)-linkedtotheglycancoreare,however,resistanttoPNGaseFwhichcanoccuronsomeplantandinsectglycoproteins.

      ToremoveO-linkedglycans,monosaccharidesmustberemovedbyaseriesofexoglycosidasesuntilonlytheGalβ1-3GalNAc(core1)and/ortheGlcNAcβ1-3GalNAc(core3)coresremainattachedtotheserineorthreonine.NEB’sO-Glycosidase,clonedfromEnterococcusfaecalis,canthenremovethesecorestructureswithnomodificationoftheserineorthreonineresidues.Anymodificationofthecorestructures,includingsialyation,willblocktheactionoftheO-Glycosidase.Sialicacidresiduesareeasilyremovedbyageneralα2-3,6,8,9NeuraminidaseA.Inaddition,exoglycosidasessuchasβ(1-4)GalactosidaseSandβ-N-Acetylhexosaminidasefcanbeincludedindeglycosylationreactionstoremoveothercomplexmodificationsoftenknowntobepresentonthecorestructures.ThiscombinationofenzymesmaynotremoveallO-linkedoligosaccharidesbutshouldremovemanycommonoligosaccharidestructures.

      TheProteinDeglycosylationMixIIcontainsalloftheenzymes,reagents,andcontrolsneededtoremoveallN-linkedandsimpleO-linkedglycansaswellassomecomplexO-linkedglycans.Thismixcontainsenzymesufficientfor20reactionsorthecleavageofasmuchas2mgofglycoprotein.AlloftheenzymesandreagentsincludedintheProteinDeglycosyationMixIIareMassSpectrometrycompatIBLe.Followingthedeglycosylationreaction,samplesarereadytobepreparedformassspectrometryanalysis.

      Figure1:AGlycoproteinmodifiedwithO-linkedandN-linkedglycosylation.

      P6044_figure_1
      Figure2


      EnzymaticDeglycosylationofBovineFetuinunderbothnative(10XDeglycosylationMixBuffer1)andreducing(10XDeglycosylationMixBuffer2)conditions.20µgreactionswereloadedontoa10-20%Tris-glycineSDS-PAGEgel.
      Lane1:   ColorPrestainedProteinStandard,BroadRange(11-245 kDa)(NEB#P7712)
      Lane2:   20 μguntreatedFetuincontrol
      Lane3:   20µgFetuindeglycosylatedundernativeconditionswithDeglycosylationMixBuffer1
      Lane4:   20µgFetuindeglycosylatedunderreducingconditionswithDeglycosylationMixBuffer2
      Lane5:   5µlProteinDeglycosylationMixII.

      ProteinDeglycosylationMixII:
      PNGaseF(Glycerol-free),Recombinant:
      10,000units/vial

      O-Glycosidase:
      80,000units/vial

      α2-3,6,8,9NeuraminidaseA:
      400units/vial

      β1-4GalactosidaseS:
      960units/vial

      β-N-acetylhexosaminidasef:
      300units/vial

      SubstrateControl:
      Fetuin,0.5mg(FetuincontainssialylatedN-linkedandO-linkedglycans)

      ofEnzymesIncludedintheProteinDeglycosylationMixII
      O-Glycosidase(NEB#P0733),alsoknownasEndo-α-N-Acetylgalactosaminidase,isarecombinantenzymeclonedfromEnterococcusfaecalis(1).Itcatalyzestheremovalofcore1andcore3O-linkeddisaccharidesfromglycoproteins.Themolecularweightisapproximately147kDa.

      PNGaseF(Glycerol-free),Recombinant(NEB#P0709),alsoknownasPeptide:N-glycosidaseF,isclonedfromElizabethkingiamiricola(formerlyFlavobacteriummeningosepticum)andexpressedinE.coli(2).PNGaseF(Glycerol-free),RecombinantisanamidasewhichcleavesbetweentheinnermostGlcNAcandasparagineresiduesofhighmannose,hybrid,andcomplexoligosaccharidesfromN-linkedglycoproteinsunlessα(1-3)corefucosylated.Themolecularweightisapproximately36kDa.

      α2-3,6,8,9NeuraminidaseA(NEB#P0722),alsoknownasSialidaseA,isarecombinantenzymeclonedfromArthrobacterureafaciensandexpressedinE.coli(3).Itcatalyzesthehydrolysisofα2,3,α2,6,α2,8andα2,9linkedN-acetylneuraminicacidresiduesfromglycoproteinsandoligosaccharides.Themolecularweightisapproximately100kDa.

      β1-4GalactosidaseS(NEB#P0745),isarecombinantenzymeclonedfromStreptococcuspneumoniaeandexpressedinE.coli(4).Itisahighlyspecificexoglycosidasethatcatalyzesthehydrolysisofβ1-4linkedgalactoseresiduesfromoligosaccharides.Themolecularweightisapproximately231kDa.

      β-N-Acetylhexosaminidasef(NEB#P0721),isarecombinantenzymeclonedfromStreptomycesplicatus(5)andoverexpressedinE.coli(6).Itcatalyzesthehydrolysisofterminalβ-N-acetylgalactosamineandglucosamineresiduesfromoligosaccharides.Themolecularweightisapproximately100kDa.

      ReagentsSupplied

      Thefollowingreagentsaresuppliedwiththisproduct:

      Storeat(°C)Concentration
      DeglycosylationMixBuffer1-2010X
      DeglycosylationMixBuffer2-2010X

      Notes:

      TheProteinDeglycosylationMixIIissuppliedwithtwodifferentreactionbuffers.Pleasereferencethe“ProtocolsandManuals”tabforthespecificprotocolassociatedwitheachbuffersystem.The10XDeglycosylationMixBuffer1(NEB#B6044)shouldbeusedwhennative(non-denaturing)conditionsarenecessary.The10XDeglycosylationMixBuffer2(NEB#B6045)willreducetheglycoprotein,butwillalsoprovidethemostefficientandcompletelevelofdeglycosylation.Ifnon-denaturingconditionsarenotrequired,werecommendusing10XDeglycosylationMixBuffer2.Storage:Itisrecommendedtostorethiskitat4°C.Allcomponentsofthekitwillbestableforatleastoneyearifstoredcorrectly.TheProteinDeglycosylationMixIIisnotrecommendedforuseonMucin-likesubstrates.
    售后保障
    蚂蚁淘生物188，秉承蚂蚁淘一贯的严谨态度，由蚂蚁淘公司专业人员负责品控、采购、物流、销售、售后，保障正品优质。以“快速好省，为科研提供好产品、好价格”为理念，直接链接原厂家，从全国各地原制造商严格挑选188款科研精品，剔除品牌溢价，188生物新电商，把好的产品带给科研！  力求给你最优质的商品。
  • Q：生物188产品正品保障吗？
    A：生物188质量把控人员具有十年的从业经验，在业界享有良好的口碑；自营商城，直接从厂家采购， 自己的团队负责国际物流和清关，中间没有第三方，所有流程严格把控，100%保证正品，假一罚十。

    Q：下单后可以修改订单吗？
    A：下单后的商品付款之前可以修改；订单付款成功，需要联系我们客服进行修改；客服电话：4000-520-616

    Q：可以开发票吗？
    A：本网站所售商品都是正规清关，均开具16%正规发票，发票金额含配送费金额，另有说明的除外。

    Q：商品几天可以发货？
    A：生物188商品，全部现货销售，付款后即可发货,一般一周内送达！

    Q：如何联系商家？
    A：有任何问题够可以电话咨询我们，全国24小时免费服务热线：4000-520-616 或联系我们的在线客服QQ：1570468124

    Q：收到的商品少了/发错了怎么办？
    A：同个订单购买多个商品可能会分为一个以上包裹发出，可能不会同时送达，建议查看订单详情是否是 部分发货状态；如未收到，可联系在线客服或者致电4000-520-616。

    Q：退换货/维修需要多长时间？
    A：一般情况下，退货处理周期为客户收到产品一个月内（以快递公司显示签收时间为准），包装规格、 数量、品种不符，外观毁损、短缺或缺陷，请在收到货24小时内申请退换货；特殊商品以合同条款为准。

客户服务

在线客服
用户反馈

何为188

极简而严谨，我们仅销售188款生物医学科研用品，款款都是爆款；因为少所以聚焦，聚焦甄选每一款产品，聚焦服务每一位客户!

关注我们 :

点击QQ联系我们
生物188微信

关注188微信公众号

获取最新优惠活动通知

  • 品质甄选，正品保证

  • 自营电商，厂家直采

  • 极简主义，188精品

常见问题 | 购买相关 | 配送与验收 | 支付问题 | 售后与服务 | 服务协议 | 隐私政策 | 何为188 | 中国智造 | 联系我们
友情链接： AnshLabs中国 | Polysciences第二代聚合物转染 | JenKem公司 | Lifespan Technologie | ATGen华东代理 | EastCoastBio | mdbiosciences授权 | 蚂蚁淘 | 微球

公司版权所有 © 2005-2021 苏州蚂蚁淘生物科技有限公司 苏ICP备17049038号-10