| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 649/665 |
| MW | N/A |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | SuperiorLabelingDyes iFluor™DyesandKits |
| Related | Generalproteins LabelingviaAminoGroups |
| Spectrum | AdvancedSpectrumViewer |
- Prepareproteinsolution(SolutionA):
Forlabeling50ugprotein(assumingthetargetproteinconcentrationis1mg/mL),mix5μL(10%ofthetotalreactionvolume)ofReactionBuffer(ComponentB)with50uLofthetargetproteinsolution.
Note1:Ifyouhaveadifferenceproteinconcentration,adjusttheproteinvolumeaccordinglytomake~50µgproteinavailableforyourlabelingreaction.
Note2:Forlabeling100ugprotein(assumingthetargetproteinconcentrationis1mg/mL),mix10uL(10%ofthetotalreactionvolume)ofReactionBuffer(ComponentB)with100uLofthetargetproteinsolution.
Note3:Theproteinshouldbedissolvedin1Xphosphatebufferedsaline(PBS),pH7.2-7.4;Iftheproteinisdissolvedinglycinebuffer,itmustbedialyzedagainst1XPBS,pH7.2-7.4,oruseAmiconUltra-0.5,Ultracel-10Membrane,10kDa(cat#UFC501008fromMillipore)toremovefreeaminesorammoniumsalts(suchasammoniumsulfateandammoniumacetate)thatarewidelyusedforproteinprecipitation.
Note4:ImpureantibodiesorantibodiesstABIlizedwithbovineserumalbumin(BSA)orgelatinwillnotbelabeledwell.
Note5:Theconjugationefficiencyissignificantlyreducediftheproteinconcentrationislessthan1mg/mL.Foroptimallabelingefficiencythefinalproteinconcentrationrangeof1-2mg/mLisrecommended. - Runconjugationreaction::
- Addtheproteinsolution(SolutionA)toONEvialoflabelingdye(ComponentA),andmixthemwellbyrepeatedlypipettingforafewtimesorvortexthevialforafewseconds.
Note:Usebothvials(ComponentA)oflabelingdyetolabel100ugproteinbydividingthe100ugproteininto2x50ugproteinandreactingeach50ugproteinwithonevialoflabelingdye.Combinetwovialsforthenextstep. - Keeptheconjugationreactionmixtureatroomtemperaturefor30-60minutes.
Note:Theconjugationreactionmixturecanberotatedorshakenforlongertimeifdesired.
- Addtheproteinsolution(SolutionA)toONEvialoflabelingdye(ComponentA),andmixthemwellbyrepeatedlypipettingforafewtimesorvortexthevialforafewseconds.
- StopConjugationreaction:
- Add5uL(for50ugprotein)or10uL(for100ugprotein)whichis10%ofthetotalreactionvolumeofTQ-DyedQuenchBuffer(ComponentC)intotheconjugationreactionmixture(fromstep2.2),mixthemwell.
- Incubateatroomtemperaturefor10minutes.
- Thelabeledprotein(antibody)isnowreadytouse.
| References&Citations |  CitationExplorer |
Cube-shapedtheranosticpaclitaxelprodrugnanocrystalswithsurfacefunctionalizationofSPCandMPEG-DSPEforimagingandchemotherapy
Authors:FuqiangGuo,JiajiaShang,HaiZhao,KangrongLai,YangLi,ZhongxiongFan,ZhenqingHou,GuanghaoSu
Journal:ColloidsandSurfacesB:Biointerfaces(2017)
Light/magnetichyperthermiatriggereddrugreleasedfrommulti-functionalThermo-sensitivemagnetoliposomesforprecisecancersynergetictheranostics
Authors:YuxinGuo,YangZhang,JinyuanMa,QiLi,YangLi,XinyiZhou,DanZhao,HuaSong,QingChen,XuanZhu
Journal:JournalofControlledRelease(2017)
Thermo-sensitivehydrogelPLGA-PEG-PLGAasavaccinedeliverysystemforintramuscularimmunization
Authors:XiaoyanWang,YuZhang,WeiXue,HongWang,XiaozhongQiu,ZonghuaLiu
Journal:JournalofBiomaterialsApplications(2017):923--932
Affinity-ControlledProteinEncapsulationintoSub-30nmTelodendrimerNanocarriersbyMultivalentandSynergisticInteractions
Authors:XuWang,ChangyingShi,LiZhang,AlexaBodman,DandanGuo,LiliWang,WalterAHall,StephanWilkens,JuntaoLuo
Journal:Biomaterials(2016)
CarboxymethylDextran-StabilizedPolyethylenimine-Poly(epsilon-caprolactone)Nanoparticles-MediatedModulationofMicroRNA-34aExpressionviaSmall-MoleculeModulatorforHepatocellularCarcinomaTherapy
Authors:XiongweiDeng,ZhaoxiaYin,ZhixiangZhou,YihuiWang,FangZhang,QinHu,YishuYang,JianqingLu,YanWu,WangSheng
Journal:ACSappliedmaterials&interfaces(2016):17068--17079
Click-electronmicroscopyforimagingmetabolicallytaggednonproteinbiomolecules
Authors:JohnTNgo,StephenRAdams,ThomasJDeerinck,DanielaBoassa,FrancesRodriguez-Rivera,SakinaFPalida,CarolynRBertozzi,MarkHEllisman,RogerYTsien
Journal:NatChemBiol(2016):459--465
Design,synthesisandevaluationofVEGF-siRNA/CRSasanovelvectorforgenedelivery
Authors:WenZhao,YifanZhang,XueyunJiang,ChunyingCui
Journal:DrugDesign,DevelopmentandTherapy(2016):3851
MolecularBasisandConsequencesoftheCytochromec-tRNAInteraction
Authors:CuipingLiu,AaronJStonestrom,ThomasChristian,JeongsikYong,RyuichiTakase,Ya-MingHou,XiaoluYang
Journal:JournalofBIOLOGicalChemistry(2016):10426--10436
Determinationoftheactivetransportoffucoidanderivedfromokinawamozukuacrossthehumanintestinalcaco-2cellsasassessedbysize-exclusionchromatography
Authors:TakeakiNagamine,KouHayakawa,KyoumiNakazato,MasahikoIha
Journal:JournalofChromatographyB(2015):187--193
Multiplexedsingle-cellinsituRNAanalysisbyreiterativehybridization
Authors:LuXiao,JiaGuo
Journal:AnalyticalMethods(2015):7290--7295
AAT Bioquest AAT Bioquest是一家位于美国的生物公司，前身为ABD Bioquest，总部位于加利福尼亚州。专门从事光学检测技术十多年，一直致力于光谱学检测领域技术的创新和突破。其独特的光学检测技术，综合了化学、生物学和信息学等各个领域的研究，引领了比色、荧光和发光技术新一代光学探针的浪潮。AAT Bioquest在全球拥有强大的经验丰富的专业分销商网络，为从小型研究机构到《财富》500强企业的各类客户提供卓越的产品和定制服务。
美国AATBioquestInc.(前身是ABDBioquest，Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域，包括吸收（颜色），荧光和发光技术。AATBioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学，免疫学，细胞生物学和分子生物学等领域。AATBioquest会不断介绍新产品，快速的丰富各个领域的产品。
1）我们提供反应荧光探针和发光探针，生物素和端粒酶能够应用于标记药物小分子和生物聚合物，如蛋白、核酸以及其他碳水化合物；2）我们研究并生产荧光和发光探针用于检测蛋白，核酸和活细胞。3）我们不断的推出新型的荧光和发光探针用于检测多种酶，特别是检测水解酶和氧化还原酶类；4）我们致力于开发用于信号转导研究的试剂；5）我们提供生理和神经探针，特别是钙离子指示剂和膜电位探针。
作为AATBioquestInc.的中国区域代理，艾美捷科技为中国客户提供光谱学检测领域，包括吸收（颜色），荧光和发光技术等全系列解决方案。我们也将一如既往更加努力为国内用户提供快捷、方便的高质量产品，同时更为您售前售后全面技术支持。
AATBioquest,Inc.(formerlyABDBioquest,Inc.)develops,manufacturesandmarketsbioanalyticalresearchreagentsandkitstoscientistsengagedinlifesciencesresearch,diagnosticR&Danddrugdiscovery.Wespecializeintheareaofphotometricdetectionsincludingabsorption(color),fluorescenceandluminescencetechnologies.TheCompany"sproductsenablescientistsandbiomedicalresearcherstobetterunderstandbiochemistry,immunology,cellBIOLOGyandmolecularbiology.AATBioquestconstantlyintroducesnewproducts,andoffersarapidlyexpandinglistofproductsthataregroupedintoseveralproductlines.
1)Ourreactivefluorescentandluminescentprobes,biotinsandtagenzymesareusedforlabelingsmalldrugmoleculesandbiopolymers,e.g,proteins,nucleicacidsandcarbohydrates;2)Wedevelopfluorescentandluminescentprobesfordetectingproteins,nucleicacidsandlivecells;3)Weconstantlyintroducenovelfluorescentandluminescentprobesfordetectingvariousenzymes,inparticular,hydrolyticandredoxenzymes;4)Wefocusondevelopingreagentsforsignaltransductionresearch;and5)Wealsoofferphysiologicalandneurologicalprobes,e.g.,calciumindicatorsandmembranepotentialprobes.
Besidesthestandardcatalogproductswealsooffercustomservicetomeetyourspecialresearchneeds.Ourcurrentservicesincludecustomsynthesisofcolorimetric,fluorescentandluminescentprobes,customdevelopmentofbiochemical,cell-basedanddiagnosticassaysandcustomscreeningofyourcompoundlibrariesagainstyourdefinedtargetsusingourvalidatedHTSassays.
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
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
