AAT Bioquest/iFluor™ 860 maleimide/1408/1 mg
市场价：
¥85560.00
美元价：
51336.00
产品分类：
荧光染料
公司分类：
Fluorescent_dyes
联系Q Q：
3392242852
电话号码：
4000-520-616
电子邮箱：
info@ebiomall.com
商品介绍
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 863/868 |
| MW | 1647.66 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | SuperiorLabelingDyes iFluor™DyesandKits |
| Related | Generalproteins LabelingviaThiolGroups |
| Spectrum | AdvancedSpectrumViewer |
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- Prepareproteinstocksolution(SolutionA):
- (Optional)ifyourproteindoesnotcontainafreecysteine,youmusttreatyourproteinwithDTTorTCEPtogenerateathiolgroup.10molarequivalentsofDTTorTCEParesufficientforconvertingadisulfidebondtotwofreethiolgroups.IfDTTisusedyoumustremovefreeDTTbydialysisorgelfiltrationbeforeconjugatingadyemaleimidetoyourprotein.Followingisasampleprotocolforgeneratingafreethiolgroup:
a.Prepareafreshsolutionof1MDTT(15.4mg/100µl)indistilledwater.
b.MakeIgGsolutionin20mMDTT:add20µlofDTTstockpermlofIgGsolutionwhilemixing.Letstandatroomtempfor30minuteswithoutadditionalmixing(tominimizereoxidationofcysteinestocystines).
c.PassthereducedIgGoverafiltrationcolumnpre-equilibratedwith"ExchangeBuffer".Collect0.25mlfractionsoffthecolumn.
d.DeterminetheproteinconcentrationsandpoolthefractionswiththemajorityoftheIgG.Thiscanbedoneeitherspectrophotometricallyorcolorimetrically.
e.Carryouttheconjugationassoonaspossibleafterthisstep(seebelow).
Note1:IgGsolutionsshouldbe>4mg/mlforthebestresults.Theantibodyshouldbeconcentratediflessthan2mg/ml.Includeanextra10%forlossesonthebufferexchangecolumn.
Note2:ThereductioncanbecarriedoutinalmostanybuffersfrompH6to7,e.g.,MES,phosphateorTRISbuffers.
Note3:Stepscanddcanbereplacedbydialysis. - Mix100µLofareactionbuffer(e.g.,100mMMESbufferwithpH~6.0)with900µLofthetargetproteinsolution(e.g.antibody,proteinconcentration>2mg/mlifpossible)togive1mLproteinlabelingstocksolution.
Note1:ThepHoftheproteinsolution(SolutionA)shouldbe6.5±0.5.
Note2:ImpureantibodiesorantibodiesstABIlizedwithbovineserumalbumin(BSA)orotherproteinswillnotbelabeledwell.
Note3:Theconjugationefficiencyissignificantlyreducediftheproteinconcentrationislessthan2mg/mL.Foroptimallabelingefficiencythefinalproteinconcentrationrangeof2-10mg/mLisrecommended.
- (Optional)ifyourproteindoesnotcontainafreecysteine,youmusttreatyourproteinwithDTTorTCEPtogenerateathiolgroup.10molarequivalentsofDTTorTCEParesufficientforconvertingadisulfidebondtotwofreethiolgroups.IfDTTisusedyoumustremovefreeDTTbydialysisorgelfiltrationbeforeconjugatingadyemaleimidetoyourprotein.Followingisasampleprotocolforgeneratingafreethiolgroup:
- Preparedyestocksolution(SolutionB):
AddanhydrousDMSOintothevialofiFluor™dyemaleimidetomakea10-20mMstocksolution.MixwellbypipettingorvortexundersuBDuedlight(ifpossible).
Note:Preparethedyestocksolution(SolutionB)beforestartingtheconjugation.Usepromptly.Extendedstorageofthedyestocksolutionmayreducethedyeactivity.SolutionBcanbestoredinfreezerforupto4weekswhenkeptfromlightandmoisture.Avoidfreeze-thawcycles. - Determinetheoptimaldye/proteinratio(optional):
Note:Eachproteinrequiresdistinctdye/proteinratio,whichalsodependsonthepropertiesofdyes.Overlabelingofaproteincoulddetrimentallyaffectsitsbindingaffinitywhiletheproteinconjugatesoflowdye/proteinratiogivesreducedsensitivity.Werecommendyouexperimentallydeterminethebestdye/proteinratiobyrepeatingSteps4and5usingaserialdifferentamountoflabelingdyesolutions.Ingeneral4-6dyes/proteinarerecommendedformostofdye-proteinconjugates.- Use10:1molarratioofSolutionB(dye)/SolutionA(protein)asthestartingpoint:Add5µlofthedyestocksolution(SolutionB,assumingthedyestocksolutionis10mM)intothevialoftheproteinsolution(95µlofSolutionA)witheffectiveshaking.Theconcentrationoftheproteinis~0.05mMassumingtheproteinconcentrationis10mg/mLandthemolecularweightoftheproteinis~200KD.
Note:TheconcentrationoftheDMSOintheproteinsolutionshouldbe<> - Runconjugationreaction(seeStep4below).
- Repeat#3.2withthemolarratiosofSolutionB/SolutionAat5:1;15:1and20:1respectively.
- Purifythedesiredconjugatesusingpremadespincolumns.
- Calculatethedye/proteinratio(DOS)fortheabove4conjugates(seenextpage).
- Runyourfunctionaltestsoftheabove4conjugatestodeterminethebestdye/proteinratiotoscaleupyourlabelingreaction.
- Use10:1molarratioofSolutionB(dye)/SolutionA(protein)asthestartingpoint:Add5µlofthedyestocksolution(SolutionB,assumingthedyestocksolutionis10mM)intothevialoftheproteinsolution(95µlofSolutionA)witheffectiveshaking.Theconcentrationoftheproteinis~0.05mMassumingtheproteinconcentrationis10mg/mLandthemolecularweightoftheproteinis~200KD.
- Runconjugationreaction:
- Addtheappropriateamountofdyestocksolution(SolutionB)intothevialoftheproteinsolution(SolutionA)witheffectiveshaking.
Note:ThebestmolarratioofSolutionB/SolutionisdeterminedfromStep3.6.IfStep3isskipped,werecommendtouse10:1molarratioofSolutionB(dye)/SolutionA(protein). - Continuetorotateorshakethereactionmixtureatroomtemperaturefor30-60minutes.
- Addtheappropriateamountofdyestocksolution(SolutionB)intothevialoftheproteinsolution(SolutionA)witheffectiveshaking.
- Purifytheconjugation:
Thefollowingprotocolisanexampleofdye-proteinconjugatepurificationbyusingaSephadexG-25column.- PrepareSephadexG-25columnaccordingtothemanufactureinstruction.
- Loadthereactionmixture(directlyfromStep4)tothetopoftheSephadexG-25column.
- AddPBS(pH7.2-7.4)assoonasthesamplerunsjustbelowthetopresinsurface.
- AddmorePBS(pH7.2-7.4)tothedesiredsampletocompletethecolumnpurification.Combinethefractionsthatcontainthedesireddye-proteinconjugate.
Note1:Forimmediateuse,thedye-proteinconjugateneedbedilutedwithstainingbuffer,andaliquotedformultipleuses.Note2:Forlongertermstorage,dye-proteinconjugatesolutionneedbeconcentratedorfreezedried(seebelow).
| References&Citations |  CitationExplorer |
Nanovesicledeliverytotheliverviaretinolbindingproteinandplatelet-derivedgrowthfactorreceptors:howtargetingligandsaffectbiodistribution
Authors:Ching-YunHsu,Chun-HanChen,IbrahimAAljuffali,You-ShanDai,Jia-YouFang
Journal:Nanomedicine(2017)
品牌介绍
美国AAT Bioquest Inc.(前身是ABD Bioquest，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。AAT Bioquest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。AAT Bioquest会不断研发新产品，快速地丰富各个领域的产品。
1）提供反应探针和发光探针，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）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）不断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）致力于开发用于信号转导研究的试剂；
5）提供生理和神经探针，特别是钙离子指示剂和膜电位探针。
品牌分类
淋巴细胞信号传导
GPCR抗体
羰基活性生物素
细胞代谢
胺反应生物素
链霉亲和素结合物
生物素化抗IgGs
乙酰基特异性抗体
细胞骨架抗体
受体结合试验
香豆素类
经典标记染料
神经酶
cAMP-GPCR分析
糖蛋白类
荧光成像
Boc氨基酸
巯基活性生物素
淬火CPG
蛋白激酶类
解理特异性抗体
胺到羧基
细胞功能分析
pH和离子指示剂
iFluor快速测试
诊断分子
氯离子通道
潮汐染料
蛋白激酶Ab
生物素结合物
金属离子
葡萄糖转运蛋白
脂肪酸转运体
MDR研究
水解酶
磷酸特异性抗体
膜电位
经典淬火剂
胺和硫醇改性剂
一般蛋白质类
胆汁酸转运蛋白
粘附力Ab
trFluor染料和试剂盒
DNA检测
β淀粉样蛋白分析
转移酵素
潮汐荧光染料和试剂盒
Fmoc氨基酸
细胞周期Ab
标记单元格
染料CPGs
细胞凋亡
菁
信号中间体Ab
荧光碳水化合物标签
酶标抗IgGs
癌症研究所
流式细胞术
肽酶和蛋白酶
报告基因酶
细胞凋亡与细胞毒性
磷酸二酯酶
活性氧种类
Cytocrhome P450酶
重要污点
染料标记试剂
染色积木
标记抗体
神经抗体
荧光抗IgGs
离子通道Ab
辣根过氧化物酶（HRP）
磷酸酶
钙GPCR测定
猝灭磷酰胺
细胞毒性
氧化还原酶
RNA检测
转录抗体
mFluor染料和试剂盒
翻译Ab
染料磷酰胺岩
iFluor染料和试剂盒
组蛋白脱乙酰酶（HDAC）
肽标记试剂
离子通道
植物蛋白
阴离子
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