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Description

The copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction between azides and alkynes to form 1,2,3-triazoles, as reported1 by Sharpless, was found to be so exquisitely regioselective and efficient at even the most mild conditions that Sharpless coined the term ‘Click Chemistry' to describe it.The use of this method for DNA modification has been somewhat delayed by the fact that copper ions damage DNA, typically yielding strand breaks.2 As these problems have now been overcome by the use of copper(I)-stabilizing ligands (e.g., tris(benzyltriazolylmethyl)amine, TBTA3), Carell et al. and Seela et al. discovered that the CuAAC reaction can be used to functionalize alkyne-modified DNA nucleobases with extremely high efficiency.4 Oligonucleotides bearing a single nucleosidic alkyne group can be prepared usinga C8-Alkyne-dC or dT-CE Phosphoramidite.Purified oligonucleotides are usually modified with 2-5 equivalents of the corresponding marker-azide (e.g., fluorescent-dye azides). After the addition of precomplexed Cu(I), complete conversion to the labelled oligo is observed in a time span between 30 min and 4 hours. After a simple precipitation step, labelled oligonucleotides can be recovered in near quantitative yields. Using a combination of C8-Alkyne, C8-TIPS-Alkyne and C8-TMS-Alkyne, it is possible to label oligonucleotides in up to three separate click reactions. The alkyne groups on the last two monomers are protected, respectively, with triisopropylsilyl (TIPS) and trimethylsilyl (TMS) protecting groups.5,6The first click reaction on solid phase on a C8-Alkyne yields the singly modified oligonucleotide with full retention of the TIPS and/or TMS protecting group.For double click, a C8-TIPS-Alkyne is used as the second nucleoside and the TIPS protecting group is cleaved with tetrabutylammonium fluoride (TBAF) without causing any damage to the DNA.The second click reaction in solution yields the doubly modified oligonucleotide in excellent yield. For the introduction of three different labels, all three nucleosides are introduced into oligonucleotides. The first click reaction is performed directly on the resin. The singly modified oligonucleotide is subsequently cleaved from the support with concomitant cleavage of the TMS group and retention of the TIPS protecting group. The second click reaction is performed in solution. Precipitation of the doubly modified oligonucleotide, cleavage of the TIPS group with TBAF, and a subsequent third click reaction in solution furnishes the desired triply modified oligonucleotide in excellent overall yield.

Details

Usage

  • Coupling: 3 minute coupling time recommended.
  • Deprotection: Ammonium hydroxide for 2 hours at 55°C or as required by nucleobases.This is more than sufficient to completely remove the TMS protecting group. Note: Deprotection in AMA (50:50 Ammonium hydroxide/MethylAmine) may result in about 5% N4-methyl-dC side product at the modification site.
Specifications
DiluentAnhydrous Acetonitrile
StorageFreezer storage, -10 to -30�C, dry
Stability1-2 days

Intellectual Property

baseclick GmbH has been granted the following patents (1-3) besides its further patent applications (4-5). |1. WO  2006/117161  (New  labelling  strategies  for  the  sensitive detection of analytes)|2. WO 2008/952775 (Click chemistry for the production of reporter molecules)|3. WO 2010/115957 (Click Chemistry on heterogeneous catalysts)|4. PCT/EP 2013/064610 (Anandamide-modified nucleic molecules)|5. PCT/EP 2015/056007 (Self-assembly of DNA Origami: a diagnostic tool)|baseclick GmbH holds a worldwide exclusive license for granted patent application|WO 03/101972 (Copper-catalysed ligation of azides and acetylenes  for  the  nucleic  acid field) in the area of diagnostics and research. As Glen Research and baseclick are partners, Glen Research is now able to help in sublicensing this outstanding technology.

Technical Documents

Safety Data Sheet

Glen Report 22.11: Simple Oligonucleotide Modification using Click Chemistry

Glen Report 22.12: The copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC)

Glen Report 23.13: New Products – Click Chemistry Update

Glen Report 24.26: Advances in copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC): Introducing Oligo-Click Kits

Glen Report 28.21: Versatile Applications of the Copper(I)-Catalyzed Click Chemistry


Dilution/Coupling Data

The table below show pack size data and, for solutions, dilution and approximate coupling based on normal priming procedures.

ABI 392/394

Catalog #Pack SizeGrams/Pack0.1M Dil. (mL)Approximate Number of Additions
LV40LV20040nm0.2μm1μm10μm
10-1542-020.25 g.25grams2.476941.425.8818.8213.83.45
10-1542-90100 µmol.101grams120127.55.4541
10-1542-9550 µmol.051grams0.53.3321.250.910.670.17

Expedite

Catalog #Pack SizeGrams/PackDilution (mL)Approximate Number of Additions
Molarity50nm0.2μm1μm15μm
10-1542-020.25 g.25grams3.690.0767.442.1330.644.21
10-1542-90100 µmol.101grams1.50.0723.614.7510.731.48
10-1542-9550 µmol.051grams0.750.078.65.383.910.54


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ICML2017:PhaseIIIbMAGNIFYStudyofLenalidomideandRituximabCombinationinRelapsed/RefractoryFollicularandMarginalZoneLymphoma

AninterimanalysisofMAGNIFY,aphaseIIIb,randomized,open-label,multicenterstudyoftheR2combinationregimen(lenalidomide[Revlimid]plusrituximab[Rituxan])inpatientswithrelapsedorrefractorymarginalzonelymphoma,waspresentedattheInternationalConferenceonMalignantLymphoma(ICML)inLugano,Switzerland.ThisanalysisexpandedupondatapresentedearlierinthemonthattheASCOAnnualMeeting(Abstract7502).

TheMAGNIFYstudycontinuestoevaluatetheclinicalactivityof12cyclesofR2combinationtherapyfollowedbyrandomizationtoeither18cyclesofR2maintenanceor18cyclesofrituximabmonotherapy,inpatientswithrelapsedorrefractoryfollicularlymphoma,marginalzonelymphoma,ormantlecelllymphoma.Approximately500patientsareplannedtobeenrolledinthestudy.

Theprimaryendpointisprogression-freesurvival.Secondaryendpointsincludeoverallsurvival,overallresponserate,completeresponse,improvementofresponse,durationofresponse,durationofcompleteresponse,timetonextlymphomatreatment,timetohistologictransformation,safety,andexploratoryquality-of-lifemeasures.EnrollmentintheMAGNIFYstudyisongoing.

ASCOData

AttheASCOAnnualMeeting,interimdatawerepresentedfromananalysisofasubsetofpatientsfromtheMAGNIFYstudywithrelapsedorrefractoryfollicularlymphoma(n=160)withearly-relapse(n=52)anddouble-refractory(n=50)disease.

AttheJanuary9,2017,datacutoff,the1-yearprogression-freesurvivalforallfollicularlymphomapatientswas70%,with65%fordouble-refractorypatientsand49%forearly-relapsepatients.Additionally,evaluablefollicularlymphomapatients(n=128)hadanoverallresponserateof66%withacompleteresponse/completeresponse–unconfirmedrateof38%.Fordouble-refractorypatients(n=42),overallresponseratewas45%withacompleteresponse/completeresponse–unconfirmedrateof21%andforearly-relapsepatients(n=43),overallresponseratewas47%withacompleteresponse/completeresponse–unconfirmedrateof21%.Mediandurationofresponsewasnotmetatamedianfollow-upof10.2months.

Themostcommongrade3or4adverseeventsobservedinthestudyforallfollicularlymphoma,double-refractory,andearly-relapsepatients,respectively,wereneutropenia(29%,42%,37%),fatigue(6%,4%,8%),leukopenia(5%,8%,10%),thrombocytopenia(4%,8%,4%),andlymphopenia(3%,6%,4%).

ICMLData

DatapresentedatICMLinaseparateanalysisfocusedonpatientswithmarginalzonelymphoma(n=38),includingnodalmarginalzonelymphoma(n=18),splenicmarginalzonelymphoma(n=10),andmucosa-associatedlymphoidtissuelymphoma(n=10).

Atamedianfollow-upof13.8monthsfrominitiationoftherapywiththeR2combination,evaluablepatientswithmarginalzonelymphoma(n=32)achievedanoverallresponserateof66%withacompleteresponse/completeresponse–unconfirmedrateof44%.Evaluablenodalmarginalzonelymphomapatients(n=14)hadanoverallresponserateof57%withacompleteresponse/completeresponse–unconfirmedrateof57%.Evaluablesplenicmarginalzonelymphomapatients(n=8)hadanoverallresponserateof63%withacompleteresponserateof25%;andevaluablemucosa-associatedlymphoidtissuelymphomapatients(n=10)hadanoverallresponserateof80%withacompleteresponse/completeresponse–unconfirmedrateof40%.Mediandurationofresponsewasnotreachedforanygroup.

Themostcommongrade3or4adverseeventsobservedinpatientswithmarginalzonelymphomawereneutropenia(32%),thrombocytopenia(16%),andleukopenia(11%).

“Thechemotherapy-freecombinationoflenalidomideandrituximab,withcomplementarymechanismsofactionthatarethoughttoenhanceantibodydependentcellularcytotoxicity,continuestoshowencouragingactivityandatolerablesafetyprofileinindolentlymphomas,andparticularlyindifficult-to-treatpatientsubsets,”saidDavidJ.Andorsky,MD,co–principalinvestigatorofthestudyandmedicaloncologistattheRockyMountainCancerCentersinBoulder,Colorado.“Theseresultsinpatientswhohadfailedmultipletherapiesorrelapsedearly,aswellastheactivityinmarginalzonepatientsmeritfurtherstudyinthisareaofindolentlymphoma.”

AboutMAGNIFY

MAGNIFYisaphaseIIIb,multicenter,open-labelstudyofpatientswithgrades1–3bortransformedfollicularlymphoma,marginalzonelymphoma,ormantlecelllymphomawhoreceivedatleast1priortherapyandhadstageI–IV,measurabledisease.Approximately500patientsareplannedforenrollmentin12cyclesofR2induction,withaprojected314patientswithatleaststablediseaseafterinductionrandomized(1:1)to2maintenancearms.

Inductionincludesorallenalidomideat20mg/d,days1–21per28-daycycle(d1–21/28)plusintravenousrituximabat375mg/m2,days1,8,15,and22ofcycle1andday1ofcycles3,5,7,9,and11(28-daycycles).Patientsarethenrandomizedtomaintenancelenalidomideat10mg/d,days1to21/28,cycles13to30,plusrituximabat375mg/m2,day1ofcycles13,15,17,19,21,23,25,27,and29(R2,armA),orrituximabalone(sameschedule,armB).PatientsreceivingR2maintenanceafter18cyclesmaycontinuemaintenancelenalidomidemonotherapyat10mg/d,days1–21/28(perpatientand/orinvestigatordiscretion),untildiseaseprogressionastolerated.Patientswillbefollowedfor≥5yearsafterthelastpatientinitiatesinductiontherapy.


名称:氨氧基乙酸盐;羧甲氧基胺半盐酸盐;

分子式:C2H5NO3·0.5HCl
分子质量:109.30
熔点:156℃

如题,之前买的sigma的粉末,太贵。加培养基又不方便

多胺的测定.123
JBDDD1232017-02-19

最近用药物处理PDX肿瘤小鼠,然后取鼠血浆,请问我是用人的ELISA,还是小鼠的Elisa呢?

还有就是我要测的是血浆中多胺的含量,Elisa有测结合态和束缚态的,这两者有什么区别,我应该如何选择呢?


果糖胺是血浆中的蛋白质在葡萄糖非酶糖化过程中形成的一种物质,由于血浆蛋白的半衰期为17天,故果糖胺反映的是1-3周内的血糖水平。
果糖胺是血浆中的蛋白质与葡萄糖非酶糖化过程中形成的高分子酮胺结构类似果糖胺的物质,它的浓度与血糖水平成正相关,并相对保持稳定。它的测定却不受血糖的影响。由于血浆蛋白的半衰期为17~20天,故果糖胺可以反映糖尿病患者检测前1~3周内的平均血糖水平。从一定程度上弥补了糖化血红蛋白不能反映较短时期内血糖浓度变化的不足。果糖胺的测定快速而价廉(化学法),是评价糖尿病控制情况的一个良好指标,尤其是对血糖波动较大的脆性糖尿病及妊娠糖尿病,了解其平均血糖水平有实际意义。但果糖胺不受每次进食的影响,所以不能用来直接指导每日胰岛素及口服降糖药的用量。血清果糖胺正常值为1.64~2.64mmol/L,血浆中果糖胺较血清低0.3mmol/L。
相关疾病:肠梗阻便秘疝气食管裂孔疝75岁老年女性,CT显示胃的位置偏高,怀疑食管裂孔疝,做造影的时候为什么选用泛葡胺而没有选用钡餐造影呢?

请问有没有大神在做,用荧光酶标仪筛选单胺氧化酶抑制剂,能不能发个具体的实验步骤啊。万分感谢!!!

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AdditionofIxazomibtoLenalidomide/DexamethasoneImprovesProgression-FreeSurvivalinRelapsed/RefractoryMultipleMyeloma

Moreauetalfoundthataddingtheoralproteasomeinhibitorixazomib(Ninlaro)tolenalidomide(Revlimid)anddexamethasonesignificantlyprolongedprogression-freesurvivalamongpatientswithrelapsed,refractory,orrelapsedandrefractorymultiplemyeloma.TheyreportedthefindingsfromthephaseIIITOURMALINE-MM1trialinTheNewEnglandJournalofMedicine.ThetrialsupportedtherecentU.S.FoodandDrugAdmiNISTration(FDA)approvalofixazomibinpreviouslytreatedmultiplemyeloma,makingitthefirstapprovedoralproteasomeinhibitor.

StudyDetails

Inthedouble-blindtrial,722patientsfrom147sitesin26countrieswererandomizedbetweenAugust2012andMay272014toreceiveixazomib(n=360)orplacebo(n=362)pluslenalidomideanddexamethasone.Treatmentconsistedof28-daycyclesoforalixazomibat4mgorplaceboondays1,8,and15;orallenalidomideat25mgondays1through21(10mginthosewithcreatinineclearance≤60or≤50mL/min/1.73m2accordingtolocalprescribinginformation);andoraldexamethasoneat40mgondays1,8,15,and22.Theprimaryendpointwasprogression-freesurvival.

Fortheixazomibvsplacebogroups,medianagewas66yearsinboth(53%vs51%>65years);58%vs56%weremale;86%vs83%werewhite;EasternCooperativeOncologyGroupperformancestatuswas0or1in95%vs93%;InternationalStagingSystem(ISS)stagewasIin63%vs64%,IIin25%vs24%,andIIIin12%inboth;78%vs72%hadcreatinineclearance≥60mL/min/1.73m2;55%vs60%hadstandard-and21%vs17%hadhigh-riskcytogenetics;numberofpriortherapieswas1in62%vs60%,2in27%vs31%,and3in11%vs9%;59%vs55%hadpriorstemcelltransplantation;diseasecategorywasrelapsedin77%inboth,refractoryin12%vs11%,relapsedandrefractoryin11%vs12%,andprimaryrefractoryin7%vs6%;priorproteasomeinhibitortherapywasbortezomib(Velcade)in69%inbothandcarfilzomib(Kyprolis)in<1%vs1%,with1%vs2%ofpatientshavingdiseaserefractorytopriortreatment;andpriorimmunomodulatorytherapywaslenalidomidein54%vs56%andthalidomide(Thalomid)in44%vs47%,with21%vs25%havingdiseaserefractorytopriortherapy.

ImprovedProgression-FreeSurvival

Aftermedianfollow-upof14.7months,medianprogression-freesurvivalwas20.6monthsintheixazomibgroupvs14.7monthsintheplacebogroup(hazardratio[HR]=0.74,P=.01).Benefitofixazomibwasconsistentacrossprespecifiedsubgroups,includingpoor-prognosissubgroupssuchasthosewithhigh-riskcytogenetics(24.1vs9.7months,HR=0.54),ISSstageIIIdisease(18.4vs10.1months,HR=0.72),thoseaged>75years(18.5vs13.1months,HR=0.87),andthosewhohadreceivedtwo(17.5vs14.1months,HR=0.75)orthree(notestimablevs10.2months,HR=0.37)priortherapies.

Overallresponserateswere78%vs72%,withtheratesofcompleteresponseplusverygoodpartialresponseof48%vs39%.Mediantimetoresponsewas1.1vs1.9months.Mediandurationofresponsewas20.5vs15.0months.Atmedianfollow-upofapproximately23months,medianoverallsurvivalhadnotbeenreachedineithergroup.

AdverseEvents

Adverseeventsof≥grade3occurredin74%oftheixazomibgroupvs69%oftheplacebogroup,withthemostcommonintheixazomibgroupbeingneutropenia(23%vs24%)andthrombocytopenia(19%vs9%).Rashofanygradewasmorecommonintheixazomibgroup(36%vs23%),asweregastrointestinaladverseevents(mostlylowgrade);22%ofixazomibpatientsand19%ofplacebopatientsreceivedantidiarrhealagents,and21%and13%receivedantiemeticdrugs.Peripheralneuropathyofanygradeoccurredin27%vs22%(grade3in2%ineach).

Seriousadverseeventsoccurredin47%vs49%.Adverseeventsledtodosereductionofanydrugin56%vs50%,discontinuationofanydrugin25%vs20%,anddiscontinuationofthestudyregimenin17%vs14%.Deathoccurredduringthestudyperiodin4%vs6%.

Theinvestigatorsconcluded:“Theadditionofixazomibtoaregimenoflenalidomideanddexamethasonewasassociatedwithsignificantlylongerprogression-freesurvival;theadditionaltoxiceffectswiththisall-oralregimenwerelimited.”



交叉配血,过程中只放1液,低离子液,离心倒去上清,摇晃,会导致红细胞凝集吗?如果凝集原理是什么?
到底有没有办法,可以把N-苄基异丙胺盐酸盐的胺味儿,彻底去除,彻底,彻底去除或者有没有办法添加某种辅料或香料,来彻底掩盖那个异胺味儿。
到底有没有办法,可以把N-苄基异丙胺盐酸盐的胺味儿,彻底去除,彻底,彻底去除或者有没有办法添加某种辅料或香料,来彻底掩盖那个异胺味儿。