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Novus/Recombinant Human TGF-beta 1 (Human Cell-expressed) Protein/25 ug/7754-BH-025
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RecombinantHumanTGF-beta1(HumanCell-expressed)ProteinSummary

DetailsofFunctionality
MeasuredbyitsABIlitytoinhibittheIL-4-dependentproliferationofHT‑2mouseT cells.Tsang, M.et al.(1995)Cytokine7:389.TheED50forthiseffectis0.04-0.2ng/mL.
Source
Humanembryonickidneycell,HEK293-derivedhumanTGF-beta1proteinAla279-Ser390
Accession#
P01137
N-terminalSequence
Ala279
Structure/Form
Disulfide-linkedhomodimer
Protein/PeptideType
RecombinantProteins
Gene
TGFB1
Purity
>95%,bySDS-PAGEunderreducingconditionsandvisualizedbysilverstain.
EndotoxinNote
<0.01 eu="" per="" 1="" μg="" of="" the="" protein="" by="" the="" lal="">

Applications/Dilutions

TheoreticalMW
12.8kDa(monomer).Disclaimernote:Theobservedmolecularweightoftheproteinmayvaryfromthelistedpredictedmolecularweightduetoposttranslationalmodifications,posttranslationcleavages,relativecharges,andotherexperimentalfactors.
SDS-PAGE
11kDa,reducingconditions
Publications
ReadPublicationsusing7754-BHinthefollowingapplications:
  • Bioassay
    4publications

Packaging,Storage&Formulations

Storage
Useamanualdefrostfreezerandavoidrepeatedfreeze-thawcycles.
  • 12monthsfromdateofreceipt,-20to-70°Cassupplied.
  • 1month,2to8°Cundersterileconditionsafterreconstitution.
  • 3months,-20to-70°Cundersterileconditionsafterreconstitution.
Buffer
Lyophilizedfroma0.2μmfilteredsolutioninAcetonitrileandTFAwithBSAasacarrierprotein.*1mgpacksize(01M)issuppliedasa0.2µmfilteredsolutioninAcetonitrileandTFAwithBSAasacarrierprotein.
Purity
>95%,bySDS-PAGEunderreducingconditionsandvisualizedbysilverstain.
ReconstitutionInstructions
Reconstituteat100μg/mLinsterile4mMHClcontainingatleast0.1%humanorbovineserumalbumin.

Notes

ThisproductisproducedbyandshipsfromR&DSystems,Inc.,aBio-Technebrand.

AlternateNamesforRecombinantHumanTGF-beta1(HumanCell-expressed)Protein

  • CEDLAP
  • DPD1
  • latency-associatedpeptide
  • TGFbeta1
  • TGFB
  • TGFB1
  • TGF-beta1protein
  • TGFbeta1
  • TGF-beta1
  • TGFbeta
  • TGF-beta-1
  • transforminggrowthfactorbeta-1
  • transforminggrowthfactor,beta1

Background

TGF‑beta1(transforminggrowthfactorbeta1)isoneofthreecloselyrelatedmammalianmembersofthelargeTGF‑betasuperfamilythatshareacharacteristiccystineknotstructure(1‑7).TGF‑beta1,‑2and‑3arehighlypleiotropiccytokinesthatareproposedtoactascellularswitchesthatregulateprocessessuchasimmunefunction,proliferationandepithelial‑mesenchymaltransition(1‑4).EachTGF‑betaisoformhassomenon‑redundantfunctions;forTGF‑beta1,micewithtargeteddeletionshowdefectsinhematopoiesisandendothelialdifferentiation,anddieofoverwhelminginflammation(2).HumanTGF‑beta1CDNAencodesa390aminoacid(aa)precursorthatcontainsa29aasignalpeptideanda361aaproprotein(8).Afurin-likeconvertaseprocessestheproproteintogenerateanN‑terminal249aalatency‑associatedpeptide(LAP)andaC‑terminal112aamatureTGF‑beta1(8,9).Disulfide-linkedhomodimersofLAPandTGF‑beta1remainnon‑covalentlyassociatedaftersecretion,formingthesmalllatentTGF‑beta1complex(8‑10).CovalentlinkageofLAPtooneofthreelatentTGF‑betabindingproteins(LTBPs)createsalargelatentcomplexthatmayinteractwiththeextracellularmatrix(9,10).TGF‑betaisactivatedfromlatencybypathwaysthatincludeactionsoftheproteaseplasmin,matrixmetalloproteases,thrombospondin1andasubsetofintegrins(10).MaturehumanTGF‑beta1shares100%aaidentitywithpig,dogandcowTGF‑beta1,and99%aaidentitywithmouse,ratandhorseTGF‑beta1.Itdemonstratescross‑speciesactivity(1).TGF‑beta1signalingbeginswithhigh‑affinitybindingtoatypeIIser/thrkinasereceptortermedTGF‑betaRII.Thisreceptorthenphosphorylatesandactivatesasecondser/thrkinasereceptor,TGF‑betaRI(alsocalledactivinreceptor‑likekinase(ALK)‑5),oralternatively,ALK‑1.ThiscomplexphosphorylatesandactivatesSmadproteinsthatregulatetranscription(3,11,12).Contributionsoftheaccessoryreceptorsbetaglycan(alsoknownasTGF‑betaRIII)andendoglin,oruseofSmad-independentsignalingpathways,allowfordisparateactionsobservedinresponsetoTGF‑betaindifferentcontexts(11).
  1. Derynck,R.andK.Miyazono(2008)ColdSpringHarborLaboratoryPressp.29.
  2. Dunker,N.andK.Krieglstein(2000)Eur.J.Biochem.267:6982.
  3. Wahl,S.M.(2006)Immunol.Rev.213:213.
  4. Chang,H.etal.(2002)Endocr.Rev.23:787.
  5. Lin,J.S.etal.(2006)Reproduction132:179.
  6. Hinck,A.P.etal.(1996)Biochemistry35:8517.
  7. Mittl,P.R.E.etal.(1996)ProteinSci.5:1261.
  8. Derynck,R.etal.(1985)Nature316:701.
  9. Miyazono,K.etal.(1988)J.Biol.Chem.263:6407.
  10. Oklu,R.andR.Hesketh(2000)Biochem.J.352:601.
  11. deCaestecker,M.etal.(2004)CytokineGrowthFactorRev.15:1.
  12. Zuniga,J.E.etal.(2005)J.Mol.Biol.354:1052.

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小弟用E.coli表达的一蛋白(一天然毒素的N段)制备了11株单抗,但没有一株能中和天然蛋白,这可急死我了.再拖下去就不能毕业了.导师天天催我建立诊断方法呢.大家给我出出主意!
有两题不解
天然蛋白质中不存在的氨基酸:同型半胱氨酸
在天然蛋白质的组成中,不含有的氨基酸是:瓜氨酸
我怎么觉得这两题一样呢
我将目的基因插入到pPIC9K中的EcoRI和NotI位点上,分泌表达得到的蛋白经信号肽切除后,在N端是不是还要比天然蛋白多8个氨基酸呀?请大家指点,谢谢。
gleason分级_123
隐幽羽2018-02-04
如题,最好有详解
最近要用原核系统表达几条基因的成熟蛋白,N末端没有Met,而且最好能够表达出可溶的有活性的蛋白。
我本想用pET带有信号肽的载体表达到周质空间,但发现在信号肽切除的位置克隆进去我的基因的话还是会引入多余的氨基酸
请教各位有经验的高手,哪些载体或者方法我可以尝试去做?
多谢了!
我想做个目的蛋白的单抗,通过基因克隆、表达、纯化来得到目的蛋白,但是因为用的是原核表达系统,缺少翻译后的正常修饰,所以肯定与天然蛋白有区别,这样的话,作出来的单抗就不一定能够识别组织中的天然蛋白了,有什么方法可以减少他们的差异?也就是尽量让单抗可以识别天然蛋白。谢谢!
http://www.medicalnewstoday.com/medicalnews.php?newsid=58498

NaturalProteinStopsDeadlyHumanBrainCancerInMice


ScientistsfromJohnsHopkinsandfromtheUniversityofMilanhaveeffectivelyproventhattheycaninhibitlethalhumanbraincancersinmiceusingaproteinthatselectivelyinducespositivechangesintheactivityofcellsthatbehavelikecancerstemcells.ThereportispublishedinNature.

Themostcommontypeofbraincancer-glioblastoma-ismarkedbythepresenceofthesestem-cell-likebraincells,which,insteadoftriggeringthereplacementofdamagedcells,formcancertissue.Stemcells,unlikeallothercellsinthebody,arecapableofformingalmostanykindofcellwhentheright"signals"triggertheirdevelopment.

Fortheirtreatmentexperiment,theresearchersreliedonaclassofproteins,bonemorphogenicproteins,thatcauseneuralstem-cell-likeclusterstolosetheirstemcellproperties,whichinturnstopstheirABIlitytodivide.

Firsttheypretreatedhumanglioblastomacellswithbonemorphogenicprotein4(BMP4),theninjectedthesetreatedcellsintomousebrains.Inmiceinjectedwithcellsthatwerenotpretreated,large,invasivecancersgrew.InthemicewithBMP4-treatedcells,nocancersgrewatall.Threetofourmonthsafterinjection,allmicethatgotuntreatedcellsdied,andnearlyallmicewithBMP4-treatedcellswerealive.

Next,thescientistsdeliveredslow-releaseBMP4-containing"beads"directlyintomousebrainswithimplantedglioblastomacells.Micethatgotemptybeadsdevelopedlargemalignanttumorsanddied.MicewithBMP4beadssurvivedmuchlonger,and80percentsurvivedfourmonthsaftercancercellimplants.

"OurideaistotreatpatientswithBMP4orsomethinglikeitrightaftersurgerytoremoveglioblastomainhopesofpreventingtheregrowthofthecancerandimprovingsurvivaltime,"saysAlessandroOlivi,M.D.,directoroftheDivisionofNeurosurgicalOncologyatHopkinsandacontributortothestudy.

OlivisaysclinicalstudiesusingBMP4couldbeginwithinayearand,ifsuccessful,drugtherapiescouldbeavailabletothepublicwithinthreetofouryears.

"ThiswasproofoftheideathatBMPscouldstopglioblastomabydepletingthestem-cell-likepopulationthatfeedsit,"saysHenryBrem,M.D.,chairmanoftheDepartmentofNeurosurgeryatHopkinsandacollaboratorinthestudy."Thisopensexcitingdoorstofutureresearchintotreatmentsandtherapiesforsuchadevastatingdisease."
如题,最近做天然蛋白WB,可是效果老不理想

所以请教一下高手
请教,天然菌种发酵的蛋白和重组技术发酵的蛋白在质量检测指标上有何区别?谢谢!
我们目前正在研究开发系列天然蛋白质降解成多肽,用于口服营养补充,目前开发的有鱼鳞胶原蛋白多肽,牦牛乳多肽等,我们在应用过程中,感觉到了他的魅力,不同原料来源、不同的工艺条件,很有很多不同的但很有意义的应用效果,非常希望能够跟战友们分享。