Description
m-Clodrosome®isamultilamellarliposomesUSPensioninwhichClodronateisencapsulatedintheaqueouscompartmentsofthemannosylatedliposomes.m-Encapsome®isformulatedandpreparedidenticallytom-Clodrosome®exceptthatclodronateisnotaddedtotheliposomes.Theliposomesarefilteredthrough2μmpolycarbonatemembranestoensurethatlargerparticles,whichmaybetoxictoanimals,areremovedfromthesuspension.Botharepreparedandpackagedundersterileconditions.Whenanimalsorcellsaretreatedwithm-Clodrosome®,phagocyticcellsrecognizetheliposomesasinvADIngforeignparticlesandproceedtoremovetheliposomesfromthelocaltissueorserumviaphagocytosis.Theliposomesthenreleaseclodronateintothecytosolresultingincelldeath.Non-encapsulatedclodronatecannotcrossthecellmembranetoinitiatecelldeath.m-Encapsome®-controlliposomes-arerecognizedandphagocytosedbythesamemechanismasm-Clodrosome®.Sincethecontrolliposomesdonotcontainclodronate,thephagocyticcellsarenotkilled.However,phagocytesdorespondtotheingestionofcontrolliposomesbycytokinesecretion,temporarysuspensionofphagocyticactivityandotherresponsesdescribedintheliterature.
Mannosereceptortargetingbymannosylatedliposomeshasbeendemonstratedforavarietyofmannosylatedlipidconjugatesinavarietyofliposomemorphologiesandcompositionsinseveraldifferentinvitroandinvivomodels.Thereareseveralpublicationsaboutusingahydrophobicderivativeofmannose(4-aminophenylalpha-D-mannopyranoside)ratherthanusingamannosylatedlipidinclodronateliposomes.Thisismainlyduetothehighcostandcomplexityofsynthesizingandconjugatingmannosetolipid.4-aminophenylalpha-D-mannopyranosideiscommerciallyavailableandfarlessexpensivethansynthesizingmannoseconjugatedlipid.
Whymannose?Mannoseisoneofthecarbohydratecomponentsofmanybacterialandviralcellsurfaces,thereforetheever-efficient,highlyredundantimmunesystemhasevolvedmultiplemechanismsforidentifyingpathogensbasedonmannoserecognition.Theanimalandplantkingdomslikewiseutilizecarbohydraterecognitionsignalingmechanismsincludingmannoseresidues.Manypublicationsevaluateothercarbohydratesastargetingmechanismsforvariouscelltypes,howevermannosetargetingtophagocytesappearstobeoneofthemorespecificmechanismsidentifiedtodate.Mammaliancellsurfaceidentificationmoleculesbasedonmannosebinding,suchastheICAMfamilyofleukocyteadhesionmolecules,targettheSIGNfamilyofmannosereceptorstoaccomplishself-recognitioninvivo.
Awell-knownandcitedstudybyUmezawa&Eto [1]demonstratesthatliposomescontainingaminophenylmannosideweremostefficientlyincorporatedintothemousebrainacrossthebloodbrainbarrier.Theradiolabeledliposomesbearingaminophenyl-alpha-D-mannopyranosideweremaximallyincorporatedintothemousebrainafter48hours,whereasinthespleenandliver,theseradioactivitiesweremaximumafter12hours.Thestudiesalsoshowedthatliposomesweremostincorporatedwasglialcellsratherthanneuronalcell.Thesubcellularfractionationstudyindicatesthatmannoselabeledliposomesareincorporatedintolysosomesrichfractionbothinliverandbrain.
Mannosylatedfluorescentliposomes(m-Fluoroliposome®)suitableformacrophagetargetingandtrackingareavailablecontainingfivedifferentfluorescentdyes(DiI,DiO,DiD,DiAandDiR)thatcoverstheentirespectrum.Fluorescentliposomescomeinstandardandmannosylatedform.Formoreinformationsee here.
Clodrosome®andEncapsome®arestandardandnon-mannosylatedreagentsthatareusedfortargetingmacrophagesinorgansandtissuesotherthancentralnervoussystems.Formoreinformationaboutthesereagentseehere.
FormulationInformation
m-Clodrosome®LiposomalClodronateSuspension
| LipidComposition | Concentration(mg/ml) | Concentration(mM) | MolarRatioPercentage |
|---|---|---|---|
| Total | 23mg/ml | 35.1mM | 100 |
| L-alpha-Phosphatidylcholine | 18.8 | 24.3 | 70 |
| Cholesterol | 4.2 | 10.9 | 30 |
| EncapsulatedDrug | Concentration |
|---|---|
| Clodronate((Dichloro-phosphono-methyl)phosphonate),DisodiumSalt | 18.4*mM |
| *Dependingonthetypeoftheclodronatesalt,itsconcentration(mg/ml)varies.Iftetrahydratesaltisused,theconcentrationoftheencapsulateddrugwillbe~7mg/ml,andifanon-hydratedsaltisused,theconcentrationwillbe~5mg/ml. | |
| Mannosylation | Concentration |
|---|---|
| 4-Aminophenyl-alpha-D-mannopyranoside | 9.53mol% |
| BufferandLiposomeSize | Specification |
|---|---|
| Buffer | PhosphateBufferedSaline |
| pH | 7.4 |
| LiposomeSize | 1.5-2µm |
TechnicalNotes
- Toreachbloodstream-accessIBLe,mannose-receptorpositivecellsoutsidetheliver,asignificantnumberofliposomeswillhavetoescapefirst-passuptakebytheliverandspleen,sothatthetargetcellsareexposedtoahigherconcentrationofmannosylatedliposomesfromtheblood.Onestrategythathasbeenusedtoensurethatliposomesescapetheliverandspleenisknownasreticuloendothelialsystem(RES)blockadeinwhichanimalsarepre-dosedwithasufficientquantityofliposomestotemporarilysaturatethephagocyticcellsoftheblood,liverandspleen,alsoknownasthereticuloendothelialsystem(RES)orthemononuclearphagocytesystem(MPS).Thissufficientquantityisdependentupontheliposometypeandcompositionaswellasthespeciesbeingdosed;thepre-dosedliposomesdonotnecessarilyneedtobethesametypeorcompositionasthetherapeuticordiagnosticliposomesavoidingtheRES.Soonafterthispre-doseisclearedfromthebloodstream(usuallywithinacoupleofhours),theliposomesofinterestaredosed.SincetheRESisinvolvedindigestingthepreviousdoseofliposomes,thesubsequentlydosedliposomeswillremaininthecirculationmuchlongerthusbemuchmorelikelytobindtotheirtargetsiteoutsidetheRESincludingthosephagocyticcellswhichareaccessible,butarenotusuallyexposedtoahigherconcentrationofliposomes.
- WhileRESblockadeisusuallythoughtofassaturatingphagocyticcells,ithasbeenshownthatopsonin-bindingbyliposomesisasaturablephenomenon.Therefore,partofRESblockademayinvolveserumdepletionofcomplementandotheropsoninsknowntocoatliposomes.Inthecurrentapplication,removalorreductionintheconcentrationofsolublemannose-receptorsmayfurtherincreasetheprobABIlityofamannosylatedliposomebeingabletointeractwithmannosereceptorsonthetargetcell.Therefore,ifthegoalistodepleteatargetsubsetofmannose-receptor+cellswhichmaynotnormallybeexposedtoasubstantialnumberofmannosylatedliposomes,pre-dosingwithmannosylatedclodronateliposomes,inordertobothsaturatetheblood,liverandspleenphagocytesandreducetheconcentrationofopsoninsincludingsolublemannosereceptors,shouldincreasethenumberofsubsequentlydosedmannosylatedclodronateliposomesavailabletothistargetsubsethypotheticallyresultinginincreaseduptakeanddepletionbythesetargetedcells.
- WhenanimalsorcellsaretreatedwithClodrosome®,phagocyticcellsrecognizetheliposomesasinvadingforeignparticlesandproceedtoremovetheliposomesfromthelocaltissueorserumviaphagocytosis.Theliposomesthenreleaseclodronateintothecytosolresultingincelldeath.Unencapsulatedclodronatecannotcrossthecellmembranetoinitiatecelldeath.
- Encapsome®controlliposomesarerecognizedandphagocytosedbythesamemechanismasClodrosome®.Sincethecontrolliposomesdonotcontainclodronate,thephagocyticcellsarenotkilled.However,phagocytesdorespondtotheingestionofthecontrolliposomesbycytokinesecretion,temporarysuspensionofphagocyticactivityandotherresponsesdescribedintheliterature.
- Theproductmustberemovedfromthevialusingsteriletechnique.Donotuseifsterilityiscompromised.Thisisparticularlyimportantifasinglevialisaccessedmultipletimesoverseveralweeks.Theproductshouldnotbeusedmorethan60daysafterreceipt,evenifunopened.
- Liposomesmaysettlewhenleftundisturbedformorethanafewhours.Immediatelypriortouse,inordertoensureahomogeneousliposomesuspension,slowlyinvertthevialseveraltimesuntilthesuspensionappearshomogeneousbyvisualinspection.Vigorousorerraticshakingwillnotdamagetheliposomesbutmayinducefoamingandbubbleformationmakingitmoredifficulttoaccuratelymeasurethedesireddosage.
- Ifthepersonnelperformingintravenousinjectionsarenotexperiencedinorfamiliarwith,precautionsforinjectinglargervolumes(~10%animalweightinml),viscousliquidsorparticulatesuspensions,considerhavingextraanimalsavailableincaseseriousinjection-relatedadverseeventsoccur.Dosecontrolanimalsfirsttobecomefamiliarwithlargevolumeinjections.
- WithinhoursaftersystemicadmiNISTrationofClodrosome®,animalsbegintoloseimportantcomponentsoftheirimmunesystem.Standardanimalhandlingandhousingprotocolsarenotsuitableforimmunocompromisedanimals.Evenwhensuchprecautionsaretaken,monitorthegeneralhealthofeachanimalforopportunisticinfectionsunrelatedtotheexperimentalprotocol.Thereisnoinherenttoxicitytotheproductattherecommendeddoselevels.
- Whendosingintravenously,usestandardprecautionsfordosinglargervolumestoanimalsincludingthefollowing:a)warmproducttoroomtemperaturepriortodosing;b)ensurethatallairbubblesareremovedfromthesyringepriortodosing.Intravenousinjectionofairbubblesmayresultinairemboliwhichcankillorseriouslyinjureanimals;c)injectproductataslow,steadyrateofnomorethan1ml/min;d)decreaseinfusionrateifanimalsdisplayanyatypicalreactionssuchasunusualagitation.
- Infusion-relatedadversereactionsusuallyinvolvetheanimalgaspingforairorotherseizure-likemovements.Animalsoftenrecoverwithnoapparentpermanentinjury,butanypotentialeffectsonexperimentalresultsmustbeassessedbytheresearcher.
- Liposomesshouldbekeptat4°CandNEVERbefrozen.
Dosage
Appearance
m-Clodrosome®andm-Encapsome®arebothwhitemilkysuspensionsmadeoflargemicrosizemultilamellarliposomes.Duetotheirlargesizesomeliposomesmightsettletothebottomofthevial.Ifleftsittingidleintherefrigerator,m-Encapsome®willphaseseparateandformpelletsinthebottomofthevialleavingaclearsolutionontop.m-Clodrosome®mightdothesameonlynotasseverely.Therefore,bothshouldbegentlyshakennottoformbubblesbuttoformahomogeneoussolutionpriortouse.
EducationalVideos
Ordering/ShippingInformation
- Allliposomebasedformulationsareshippedonblueiceat4°Cininsulatedpackagesusingovernightshippingorinternationalexpressshipping.
- LiposomesshouldNEVERbefrozen.Icecrystalsthatforminthelipidmembranecanrupturethemembrane,changethesizeoftheliposomesandcausetheencapsulateddrugtoleakout.Liposomesinliquidformshouldalwaysbekeptintherefrigerator.
- ClientswhoorderfromoutsideoftheUnitedStatesofAmericaareresponsiblefortheirgovernmentimporttaxesandcustomspaperwork.EncapsulaNanoSciencesisNOTresponsibleforimportationfeestocountriesoutsideoftheUnitedStatesofAmerica.
- WestronglyencouragetheclientsinJapan,Korea,TaiwanandChinatoorderviaadistributor.Toughcustomsclearanceregulationsinthesecountrieswillcausedelayincustomclearanceoftheseperishableformulationsifordereddirectlythroughus.Distributorscaneasilyclearthepackagesfromcustoms.Toseethelistofthedistributorsclickhere.
- ClientsorderingfromuniversitiesandresearchinstitutesinAustraliashouldkeepinmindthattheliposomeformulationsaremadefromsyntheticmaterialandtheformulationsdonotrequirea“permittoimportquarantinematerial”.LiposomesareNOTBIOLOGicalproducts.
- Ifyouwouldlikeyourinstitute’sFedExorDHLaccounttobechargedforshipping,thenpleaseprovidetheaccountnumberatthetimeofordering.
- EncapsulaNanoScienceshasnocontroloverdelaysduetoinclementweatherorcustomsclearancedelays.YouwillreceiveaFedExorDHLtrackingnumberonceyourorderisconfirmed.ContactFedExorDHLinadvanceandmakesurethatthepaperworkforcustomsisdoneontime.AllsubsequentshippinginquiriesshouldbedirectedtoFederalExpressorDHL.
StorageandShelfLife
Storage
m-Clodrosome®andm-Encapsome®shouldalwaysbestoredatinthedarkat4°C,exceptwhenbroughttoroomtemperatureforbriefperiodspriortoanimaldosing.DONOTFREEZE.Ifthesuspensionisfrozen,clodronatecanbereleasedfromtheliposomesthuslimitingitseffectivenessindepletingmacrophages.ENSisnotresponsibleforresultsgeneratedbyfrozenproduct.
ShelfLife
m-Clodrosome®andm-Encapsome®aremadeondailybasis.Thebatchthatisshippedismanufacturedonthesameday.Itisadvisedtousetheproductswithin60daysofthemanufacturingdate.
Referencesandbackgroundreading
1.UmezawaFA,EtoY.Liposometargetingtomousebrain:mannoseasarecognitionMarker.Biochemicalandbiophysicalresearchcommunications.1988Jun30;153(3):1038-44.
2.ItoT,IshigamiM,MatsushitaY,HirataM,MatsubaraK,IshikawaT,HibiH,UedaM,HirookaY,GotoH,YamamotoA.SecretedEctodomainofSIGLEC-9andMCP-1SynergisticallyImproveAcuteLiverFailureinRatsbyAlteringMacrophagePolarity.Scientificreports.2017Mar8;7:44043.
3.MironVE,BoydA,ZhaoJW,YuenTJ,RuckhJM,ShadrachJL,vanWijngaardenP,WagersAJ,WilliamsA,FranklinRJ.M2microgliaandmacrophagesdriveoligodendrocytedifferentiationduringCNSremyelination.Natureneuroscience.2013Sep;16(9):1211.
4.AndreouK,SarmientoSotoM,AllenD,EconomopoulosV,deBernardiA,LarkinJ,SibsonNR.Anti-InflammatoryMicroglia/MacrophagesasaPotentialTherapeuticTargetinBrainMetastasis.Frontiersinoncology.2017;7:251.
5.AlishekevitzD,Gingis-VelitskiS,Kaidar-PersonO,Gutter-KaponL,SchererSD,RavivZ,MerquiolE,Ben-NunY,MillerV,Rachman-TzemahC,TimanerM.Macrophage-inducedlymphangiogenesisandmetastasisfollowingpaclitaxelchemotherapyisregulatedbyVEGFR3.Cellreports.2016Oct25;17(5):1344-56.
6.OhSH,KimHN,ParkHJ,ShinJY,BaeEJ,SunwooMK,LeeSJ,LeePH.Mesenchymalstemcellsinhibittransmissionofalpha-synucleinbymodulatingclathrin-mediatedendocytosisinaParkinsonianmodel.Cellreports.2016Feb2;14(4):835-49.
7.KanoF,MatsubaraK,UedaM,HibiH,YamamotoA.SecretedEctodomainofSialicAcid‐BindingIg‐LikeLectin‐9andMonocyteChemoattractantProtein‐1SynergisticallyRegenerateTransectedRatPeripheralNervesbyAlteringMacrophagePolarity.STEMCELLS.2017Mar1;35(3):641-53.
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将来可能会出现基于免疫方法的试剂盒，不过因为H7N9是个新病毒，爆发至今不足一月，这么快时间还来不及生产抗体，所以目前的检测试剂盒只能是PCR检测试剂盒。
查到了：
4月7日，上海之江生物科技有限公司官方微博称，该公司成功研制禽流感H7N9(2013)核酸测定试剂盒(荧光PCR法)，是针对国内此次H7N9病毒最早研制成功的产品，也是国内目前唯一供应的成品化试剂盒。
从1～2个T75培养皿的感染细胞培养液中纯化腺病毒
产品简介
本系列试剂盒可快速，高效地从腺病毒感染的细胞培养液中分离纯化腺病毒，相比于传统CsCl超速离心的腺病毒病毒纯化方式（需要24个小时才能完成纯化），本试剂盒可以在1个小时内完成病毒纯化，操作简单，快速，纯化率高，纯化到的病毒颗粒可直接用于下游实验，如细胞和动物感染。
产品特点
?操作简单，快速，可以在1个小时内完成病毒纯化，不依赖于超速离心操作。
?纯化效率高：小量纯化，可从1~2个T75瓶培养的细胞培养液中纯化病毒颗粒高达1x1012VPs
?每个纯化柱，可以重复利用一次，用于纯化相同种类的腺病毒。
保存条件
纯化柱和脱盐柱保存于4℃，其它组分室温保存。
试剂盒组分
Catalog#
V1160-01
Notes
Preps
10
MiniColumns
5
Canbeusedtwice
(Storeat4°C)
Press-OnCap
5
StoreatRT
DesaltingTube*
1
Canberegenarated
(Storeat4°C)
15mLCollectionTube
10
StoreatRT
10xWashBuffer
30mL
StoreatRT
2xElutionBuffer
30mL
StoreatRT
RegenerationBuffer
30mL
StoreatRT
联系方式：
Biomiga(中国）
电话：0573-82651206
QQ:441931287
联系人：张小姐
如果PCR产物不是很纯，或者PCR扩增条带比较小，PCR产物前面又有较多引物二聚体时，用胶回收，其余用PCR产物纯化试剂盒。
pcr纯化试剂盒和胶回收试剂盒的区别：
PCR纯化试剂盒：是直接水溶解的PAC产物就可以回收，回收效率高，但是只适合单一条带需要纯化测序的时候使用。
PCR凝胶试剂盒：是在PCR产物是混合物，有多条杂带的情况下，先跑胶将杂带分离，然后在将所要的条带位置的胶切下回收，后者的回收效率低，但是很纯净。
胶回收试剂盒操作步骤：
配制琼脂糖EB凝胶,电泳以分离DNA片段。任何类型或等级的琼脂糖都可以使用。
电泳足够时间后，在紫外灯下小心地把所需的DNA的片段切下来。并尽量去除多余的凝胶。
称取空离心管的重量，切下带目的片段的凝胶装在1.5ml离心管中并称其重量，求出凝胶块的重量，近似地确定其体积。一般情况下，凝胶的密度为1g/ml，于是凝胶的体积与重量的关系可按下面换算：凝胶薄片的重量为0.2g 则其体积为0.2ml；加入等倍凝胶体积的Binding Buffer，把混合物置于55℃~65℃水浴中温浴7min至凝胶完全融化，其间每隔2-3分钟混匀一次；
转移700μl的DNA-琼脂糖溶液到一个HiBindTM DNA柱子，并把柱子装在一个干净的2ml收集管内，室温下，10,000×g离心1min，弃去液体。
将柱子重新套回收集管中，加300μl Binding Buffer至HiBind DNA 柱子中；室温下，10,000×g离心 1分钟，去弃滤出液；这一步相当关键，不要忽略此步。
将柱子重新套回收集管中，加入700μl SPW Wash buffer至HiBind DNA柱子中，室温下，10,000×g离心1分钟，去弃滤出液；注：SPW Wash buffer在使用前必须按瓶子标鉴要求用无水乙醇进行稀释。
将柱子重新套回收集管中，重复加入700μl SPW Wash buffer至HiBind DNA柱子中，室温下，10,000×g离心1分钟，去弃滤出液；
弃去液体，将空柱子重新套回收集管中，10,000×g离心1min以甩干柱基质残余的液体。
这步可以去除柱子基质上残余的乙醇，不要省略此步―――对得到好的DNA产量是十分重要的。
把柱子装在一个干净的1.5ml离心管上，加入30~50μl洗脱液或灭菌水上柱子膜上，10,000×g离心1分钟，离心管中的溶液就是纯化的DNA产物，保存于-20度。
短截就是把枝条剪短，主要作用是促使其抽生新梢，增加分枝数目，以保证树势健状和正常结果。短截常用于骨干枝组修剪，结果枝组修剪，和树体局部更新复状。
短截按其长度可分为：
① 中短截：在一年生枝的中部短截，剪后萌发的顶端枝条，长势强，下部枝条长势弱。
② 重短截：剪去一年生枝的2/3。剪后萌发出的枝条较强状，一般用于主侧枝延长头修剪。
③ 重剪：剪去一年生枝的3/4－4/5，剪后萌发出的枝条长势强状，常用于发育枝作延长枝头和徒长果枝，中果枝的修剪。
④极重短截：剪去一年生枝的4/5以上，萌发后的枝条中庸偏状，常用于将发育枝和徒长枝培养结果枝组。
⑤留基部2芽剪：剪后萌发枝条较旺盛，常用于预备枝的修剪。对于幼龄树，树势较旺，以培养良好而牢固的树形结构，提早结果为主要目的，以轻短截，少疏间为主，从始果期到盛果期，主要使桃树多结果，并形成好的树形。
如果用组织DNA提取试剂盒，提出来的就是组织细胞的DNA了
