
Description
Clodrosome®isamultilamellarliposomesUSPensioninwhichClodronateisencapsulatedintheaqueouscompartmentsoftheliposomes.Encapsome®isformulatedandpreparedidenticallytoClodrosome®exceptthatclodronateisnotaddedtotheliposomes.Theliposomesarefilteredthrough2μmpolycarbonatemembranestoensurethatthelargerparticles,whichmaybetoxictoanimals,areremovedfromthesuspension.Botharepreparedandpackagedundersterileconditions.WhenanimalsorcellsaretreatedwithClodrosome®,phagocyticcellsrecognizetheliposomesasinvADIngforeignparticlesandproceedtoremovetheliposomesfromthelocaltissueorserumviaphagocytosis.Theliposomesthenreleaseclodronateintothecytosol,resultingincelldeath.Non-encapsulatedclodronatecannotcrossthecellmembranetoinitiatecelldeath.
Controlliposomes(Encapsome®)arerecognizedandphagocytosedbythesamemechanismasClodrosome®.Sincethecontrolliposomesdonotcontainclodronate,thephagocyticcellsarenotkilled.However,phagocytesdorespondtotheingestionofcontrolliposomesbycytokinesecretion,temporarysuspensionofphagocyticactivityandotherresponsesdescribedintheliterature.
MacrophagedepletionkitiscomposedofonevialofClodrosome®andonevialof Encapsome®.Asanexample,a5-mlkitiscomposedof 5mlClodrosome®and5mlEncapsome®.
m-Clodrosome®andm-Encapsome®aremannosylatedreagentsthatarespecificallyformulatedtoefficientlytargetthemacrophagesincentralnervoussystemsandmacrophagesthatcontainmoremannosereceptors.Formoreinformationaboutthesereagentseehere.
Fluorescentliposomes(Fluoroliposome®)suitableformacrophagetargetingandtrackingareavailablecontainingfivedifferentfluorescentdyes(DiI,DiO,DiD,DiAandDiR)thatcoverstheentirespectrum.Fluorescentliposomescomeinstandardandmannosylatedform.Formoreinformationsee here.
TechnicalInformation
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. |
Encapsome®ControlLiposomeSuspension
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 |
BufferandLiposomeSize | Specification |
---|---|
Buffer | PhosphateBufferedSaline |
pH | 7.4 |
LiposomeSize | 1.5-2µm |
Dosage
TechnicalNotes
- 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;decreaseinfusionrateifanimalsdisplayanyatypicalreactionssuchasunusualagitation.
- Infusion-relatedadversereactionsusuallyinvolvetheanimalgaspingforairorotherseizure-likemovements.Animalsoftenrecoverwithnoapparentpermanentinjury,butanypotentialeffectsonexperimentalresultsmustbeassessedbytheresearcher.
- Liposomesshouldbekeptat4°CandNEVERbefrozen.
Appearance
Clodrosome®andEncapsome®arebothwhitemilkysuspensionsmadeoflargemicrosizemultilamellarliposomes.Duetotheirlargesize,someliposomesmightsettletothebottomofthevial.Ifleftsittingidleintherefrigerator,Encapsome®willphaseseparateandformpelletsinthebottomofthevialleavingaclearsolutionontop.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
Clodrosome®andEncapsome®shouldalwaysbestoredatinthedarkat4°C,exceptwhenbroughttoroomtemperatureforbriefperiodspriortoanimaldosing.DONOTFREEZE.Ifthesuspensionisfrozen,clodronatecanbereleasedfromtheliposomesthuslimitingitseffectivenessindepletingmacrophages.ENSisnotresponsibleforresultsgeneratedbyfrozenproduct.
ShelfLife
Clodrosome®andEncapsome®aremadeondailybasis.Thebatchthatisshippedismanufacturedonthesameday.Itisadvisedtousetheproductswithin60daysofthemanufacturingdate.
Referencesandbackgroundreading
1.HinsonSR,CliftIC,LuoN,KryzerTJ,LennonVA.Autoantibody-inducedinternalizationofCNSAQP4waterchannelandEAAT2glutamatetransporterrequiresastrocyticFcreceptor.ProceedingsoftheNationalAcademyofSciences.2017May23;114(21):5491-6.
2.DhupkarP,GordonN,StewartJ,KleinermanES.Anti‐PD‐1therapyredirectsmacrophagesfromanM2toanM1phenotypeinducingregressionofOSlungmetastases.CancerMedicine.2018May7.
3.XiongY,PageJC,NarayananN,WangC,JiaZ,YueF,ShiX,JinW,HuK,DengM,ShiR.Peripheralneuropathyandhindlimbparalysisinamousemodelofadipocyte-specificknockoutofLkb1.EBioMedicine.2017Oct1;24:127-36.
4.CriderA,FengT,PandyaCD,DavisT,NairA,AhmedAO,BabanB,TureckiG,PillaiA.Complementcomponent3areceptordeficiencyattenuateschronicstress-inducedmonocyteinfiltrationanddepressive-likebehavior.Brain,behavior,andimmunity.2018Mar5.
5.KocherT,AsslaberD,ZaborskyN,FlenadyS,DenkU,ReinthalerP,AblingerM,GeisbergerR,BauerJW,SeiffertM,HartmannTN.CD4+Tcells,butnotnon-classicalmonocytes,aredispensableforthedevelopmentofchroniclymphocyticleukemiaintheTCL1-tgmurinemodel.Leukemia.2016Jun;30(6):1409.
6.ZhuZ,DingJ,MaZ,IwashinaT,TredgetEE.Systemicdepletionofmacrophagesinthesubacutephaseofwoundhealingreduceshypertrophicscarformation.WoundRepairandRegeneration.2016Jul1;24(4):644-56.
7.HaqueMR,LeeDY,AhnCH,JeongJH,ByunY.Localco-deliveryofpancreaticisletsandliposomalclodronateusinginjectablehydrogeltopreventacuteimmunereactionsinatype1diabetes.Pharmaceuticalresearch.2014Sep1;31(9):2453-62.
8.MayoL,CunhaAP,MadiA,BeynonV,YangZ,AlvarezJI,PratA,SobelRA,KobzikL,LassmannH,QuintanaFJ.IL-10-dependentTr1cellsattenuateastrocyteactivationandamelioratechroniccentralnervoussysteminflammation.Brain.2016May31;139(7):1939-57.
9.KermanizadehA,ChauchéC,BalharryD,BrownDM,KanaseN,BoczkowskiJ,LanoneS,StoneV.TheroleofKupffercellsinthehepaticresponsetosilvernanoparticles.Nanotoxicology.2014Aug31;8(sup1):149-54.
10.NandiB,ShapiroM,SamurMK,PaiC,FrankNY,YoonC,PrabhalaRH,MunshiNC,GoldJS.StromalCCR6drivestumorgrowthinamurinetransplantablecoloncancerthroughrecruitmentoftumor-promotingmacrophages.Oncoimmunology.2016Aug2;5(8):e1189052.
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一、免疫组化试剂盒一般包括:
1、特异性的一抗
2、免疫组化检测系统。
3、有的同时还具备显色系统。
二、不同的公司内容有差别,所以购买时一定了解清楚:
1、一抗是做人的还是兔、鼠的组织;
2、检测系统是ABC/SP/非生物素/其他;
3、试剂盒的内容都有什么?有没有显色系统?
4、如果没有,自己根据前面的检测系统需配哪个显色系统。(其他的辅助试剂如PBS缓冲液、抗原修复液等一般另卖或自配)
三、免疫组化抗体即指特异性一抗,是用来标记的指标,免疫组化过程的其他试剂一律另外配备。
另外:做实验时,如遇出结果困难,记得做阳性片对照,敢于怀疑检测系统或显色系统的问题。
用双氧水去除过氧化物酶时,我是自己配的3%双氧水滴到片上而非将片浸入双氧水,这样可否?
用博士德的DAB染色,配制方法为:1mlH20中加入DAB、H2O2、TBS各一滴,我配制的时候发现将DAB加入H20时很容易沉淀,要震荡才能混匀,且将配好的DAB滴到玻片上时,DAB会结为很小块的微颗粒悬浮在液体中但并不染色,是否DAB有问题?
现在打算换中衫的试剂盒,代理商说中衫的试剂盒包括了二抗和DAB,想请教一下该试剂盒的具体内容有些什么?
万分感谢!
上海好多的供应商的,具体还是看你选择了,其实试剂盒产品都差不多,什么远慕生物,古朵生物...好多的额