Description
ThekitiscomposedofonevialofPEGylateddoxorubicinliposomesandonevialofPEGylatedcontrolliposomes. PEGylateddoxorubicinliposomeformulationissimilar tocommercialDoxil®insize,lipidcompositionandtheamountofencapsulateddrug.Theformulationisforresearchpurposesonlyandforinjectiontolaboratoryanimals.DoxorubicindrugisalreadyloadedintotheliposomesusingremoteloADIngbyammoniumsulfategradient.Theformulationisreadyforinjection. ThecontrolformulationofPEGylateddoxorubicinissimilartoPEGylateddoxorubicininsizeandlipidcomposition,butitdoesnotcontaindoxorubicindrug.Theformulationismadeinammoniumsulfategradient.
Surfacereactiveliposomeformulationssuitableforconjugationtovariousantibodies,proteins,peptidesandligandsarealsoavailable.Formoreinformationaboutsurfacereactiveliposomes(Immunsome®)seehere.
Formoreinformationaboutsurfacereactivedoxorubicinliposomes(Immundox®)suitableforconjugationseehere.
FormulationInformation
StandardDoxosome™Kit-DoxorubicinLiposomes(PEGylated)
| 2-VialKitforStandardDoxosome™(PEGylated) | Specification |
|---|---|
| Vial1 | PreformedliposomescomposedofHSPC:Cholesterol:DSPE-PEG(2000)(57:38:5molarratio) |
| Vial2 | PreformedliposomescomposedofHSPC:Cholesterol:DSPE-PEG(2000)(57:38:5molarratio) |
| LipidCompositionforVial1 | Concentration(mg/ml) | Concentration(mM) | MolarRatioPercentage |
|---|---|---|---|
| Total | 15.96mg/ml | 21.62mM | 100 |
| HydrogenatedSoyPC | 9.58 | 12.22 | 57 |
| Cholesterol | 3.19 | 1.14 | 38 |
| DSPE-PEG(2000) | 3.19 | 8.26 | 5 |
| Buffers,LiposomeSizeandEncapsulatedDrugConcentrationforVial1 | Specification |
|---|---|
| InsideBuffer | AmmoniumSulfate |
| OutsideBuffer | Histidine/SucroseBuffer |
| pH | 6.5 |
| LiposomeSize | 80-100nm |
| EncapsulatedDoxorubicin | 2mg/ml(3.45mM) |
| LipidCompositionforVial2 | Concentration(mg/ml) | Concentration(mM) | MolarRatioPercentage |
|---|---|---|---|
| Total | 15.96mg/ml | 21.62mM | 100 |
| HydrogenatedSoyPC | 9.58 | 12.22 | 57 |
| Cholesterol | 3.19 | 1.14 | 38 |
| DSPE-PEG(2000) | 3.19 | 8.26 | 5 |
| BuffersandLiposomeSizeforVial2 | Specification |
|---|---|
| InsideBuffer | AmmoniumSulfate |
| OutsideBuffer | Histidine/SucroseBuffer |
| pH | 6.5 |
| LiposomeSize | 80-100nm |
LiposomeParticleCalculator
Doxorubicinliposomesareunilamellarandsizedto100nm.Themolarconcentrationofliposomeis21.62mM.Byhaving liposomediameter(nm)andlipidconcentration(µM),youcancalculatethetotalnumberofthelipidsinoneliposomeandthenumberoftheliposomesinonemilliliteroftheliposomesolution.Tousethecalculatorclick here.
Appearance
Doxosome™isared,andDoxosome™Controlisawhitetranslucentliquidbothmadeofnanosizeunilamellarliposomes.Usually,duetothesmallsizeofliposomes,nosettlingwilloccurinthebottomofthevial.Theliposomesarepackagedinambervials.
EducationalVideo
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
Doxosome™productsshouldalwaysbestoredatinthedarkat4°C,exceptwhenbroughttoroomtemperatureforbriefperiodspriortoanimaldosing.DONOTFREEZE.IfthesUSPensionisfrozen,theencapsulateddrugcanbereleasedfromtheliposomesthuslimitingitseffectiveness.Inaddition,thesizeoftheliposomeswillalsochangeuponfreezingandthawing.
ShelfLife
Doxosome™ismadeondailybasis.Thebatchthatisshippedismanufacturedonthesameday.Itisadvisedtousetheproductswithin4monthsofthemanufacturingdate.
Referencesandbackgroundreading
1.HermansonGT.Bioconjugatetechniques.Academicpress;2013Jul25.
2.TorchilinV,WeissigV,editors.Liposomes:apracticalapproach.OxfordUniversityPress;2003Jun5.
3.GrabarekZ,GergelyJ.Zero-lengthcrosslinkingprocedurewiththeuseofactiveesters.Analyticalbiochemistry.1990Feb15;185(1):131-5.
4.YanL,CraytonSH,ThawaniJP,AmirshaghaghiA,TsourkasA,ChengZ.ApH‐ResponsiveDrug‐DeliveryPlatformBasedonGlycolChitosan–CoatedLiposomes.Small.2015Oct1;11(37):4870-4.
5. Silva-LópezEI,EdensLE,BardenAO,KellerDJ,BrozikJA.ConditionsforliposomeadsorptionandbilayerformationonBSApassivatedsolidsupports.Chemistryandphysicsoflipids.2014Oct31;183:91-9.
6.HazraM,SinghSK,andRayS.SurfaceModificationofLiposomalVaccinesbyPeptideConjugation.JournalofPharmaSciTech,2011;1(1):41-47.
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其检测原理为：在正常的活细胞中，磷脂酰丝氨酸（phosphotidylserine，PS）位于细胞膜的内侧，但在早期凋亡的细胞中，PS 从细胞膜的内侧翻转到细胞膜的表面，暴露在细胞外环境中。Annexin-Ⅴ（膜联蛋白-V）是一种分子量为35-36KD的Ca2+ 依赖性磷脂结合蛋白，能与PS高亲和力结合。可通过细胞外侧暴露的磷脂酰丝氨酸与凋亡早期细胞的胞膜结合。
操作步骤：
1.从室温平衡20min后的铝箔袋中取出所需板条，剩余板条用自封袋密封放回4℃。
2.设置标准品孔和样本孔，标准品孔各加不同浓度的标准品50μL；
3.样本孔中加入待测样本50μL；空白孔不加。
4.除空白孔外，标准品孔和样本孔中每孔加入辣根过氧化物酶（HRP）标记的检测抗体100μL，用封板膜封住反应孔，37℃水浴锅或恒温箱温育60min。
5.弃去液体，吸水纸上拍干，每孔加满洗涤液（350μL），静置1min，甩去洗涤液，吸水纸上拍干，如此重复洗板5次（也可用洗板机洗板）。
6.每孔加入底物A、B各50μL，37℃避光孵育15min。
7.每孔加入终止液50μL，15min内，在450nm波长处测定各孔的OD值。
不知道发在这里合适不，实在是求助无门啊！版主手下留情。
想请问下那个公司有专门的蛋白质荧光标记试剂盒出售。最好的是CY5的，我准备做三标。性价比越高越好
那些做过的前辈们指导一下。
2、 血浆：EDTA、柠檬酸盐、肝素血浆可用于检测。1000×g离心30分钟去除颗粒。
3、 细胞上清液：1000×g离心10分钟去除颗粒和聚合物。
4、 组织匀浆：将组织加入适量生理盐水捣碎。1000×g离心10分钟，取上清液。
5、 保存：如果样品不立即使用，应将其分成小部分-70℃保存，避免反复冷冻。尽可能的不要使用溶血或高血脂血。如果血清中大量颗粒，检测前先离心或过滤。不要在37℃或更高的温度加热解冻。应在室温下解冻并确保样品均匀地充分解冻。
此IBL试剂盒能用于小鼠血清,EDTA血浆,细胞上清中白介素-6的定量检测 试剂盒成分 1 预包被板: 抗小鼠白介素-6兔子IgG,亲合纯化 96T 2 酶标记抗体: (30倍浓缩)HRP标记抗小鼠白介素-6兔子IgG,亲合纯化 0.4mL x 1 3 标准品: 重组小鼠白介素-6 0.5mL x 2 4 EIA缓冲液: 含1% BSA, 0.05%吐温20 BPS 30mL x 1 5 标记抗体稀释液: 含1% BSA, 0.05%吐温20 BPS 12mL x 1 6 显色剂: TMB底物液 15mL x 1 7 终止液: 1N硫酸 12mL x 1 8 浓缩洗涤液: (40倍浓缩) 含1% BSA, 0.05%吐温20 BPS 50mL x 1 操作说明 1实验所需器材(但试剂盒没有提供) 酶标仪(450nm) 微移液管及其吸嘴 量筒及烧杯 去离子水 冰箱(4°C) 坐标纸(log/log) 吸水纸 试管(用于标准品稀释) 温育箱(37°C ± 1°C) 洗瓶 (用于洗板) 一次性试剂管(用于浓缩酶标记抗体和显色剂)向左转|向右转
