Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 360/449 |
MW | 256.15 |
CAS# | 3368-04-5 |
Solvent | Water |
Storage | F/D/L |
Category | EnzymeDetection Phosphatases |
Related | ProteinPhosphatase BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
1.Prepareworkingsolution:
1.1 Preparea2to10mMstocksolutioninanappreciatesolvent(seeTable).Thestocksolutionshouldbeusedpromptly.Anyunusedsolutionneedtobealiquotedandfrozenat<-20oC.
Note1:DonotuseDMSO,ETOHorMETHtomakestocksolutionofSunRed™Phosphate(cat#11629)sinceitsignificantlyincreasesassaybackground.
Note2:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.
1.2 Preparea2XPhosphateworkingsolution: Onthedayoftheexperiment,eitherdissolvethesubstrateinanappreciatesolventorthawanaliquotofthestocksolution(fromStep1.1)atroomtemperature.Preparea2Xworkingsolutionof10to50µMin100mMTrisbufferorbufferofyourchoice,pH8to9(notphosphatebuffer).
2.Runphosphataseassayinsupernatants:
2.1 Add50µLof2XPhosphateworkingsolution(fromStep1.2)intoeachwellofthephosphatasestandard,blankcontrol,andtestsamplestomakethetotalphosphataseassayvolumeof100µL/well.
Note:Fora384-wellplate,add25µLofsampleand25µLof2XPhosphateworkingsolutionintoeachwell.
2.2 Incubatethereactionfor30to120minutesatthedesiredtemperature,protectedfromlight.
2.3 Monitorthefluorescenceincreaseatanappropriatefiltersets(Seetable)withafluorescenceplatereader.
2.4 Thefluorescenceinblankwells(withtheassaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosewellswiththephosphatasereactions.
References&Citations | PrinterFriendlyVersion |
1. BednarskaK,KlinkM,SulowskaZ.(2006)Applicationofintracellularalkalinephosphataseactivitymeasurementindetectionofneutrophiladherenceinvitro.MediatorsInflamm,2006,19307.
2. WohlsenT,BaylissJ,GrayB,BatesJ,KatouliM.(2006)EvaluationofanalternativemethodfortheenumerationandconfirmationofClostridiumperfringensfromtreatedanduntreatedsewages.LettApplMicrobiol,42,438.
3. AndersonBH,MagdoffFR.(2005)Relativemovementandsoilfixationofsolubleorganicandinorganicphosphorus.JEnvironQual,34,2228.
4. RinconV,CorredorA,Martinez-GutierrezM,CastellanosJE.(2005)Fluorometriccell-ELISAforquantifyingrABIesinfectionandheparininhibition.JVirolMethods,127,33.
5. HoehamerCF,MazurCS,WolfeNL.(2005)PurificationandpartialcharacterizationofanacidphosphatasefromSpirodelaoligorrhizaanditsaffinityforselectedorganophosphatepesticides.JAgricFoodChem,53,90.
6. ShroyerML,BhuniaAK.(2003)Developmentofarapid1-hfluorescence-basedcytotoxicityassayforListeriaspecies.JMicrobiolMethods,55,35.
7. ChruszczM,LaidlerP,MonkiewiczM,OrtlundE,LebiodaL,LewinskiK.(2003)CrystalstructureofacovalentintermediateofendogenoushumanarylsulfataseA.JInorgBiochem,96,386.
8. BautersTG,PelemanR,DhontM,VanhaesebrouckP,NelisHJ.(2003)EnzymaticdifferentiationofCandidaparapsilosisfromotherCandidaspp.inamembranefiltrationtest.JMicrobiolMethods,53,11.
9. BouaichaN,MaatoukI,VincentG,LeviY.(2002)Acolorimetricandfluorometricmicroplateassayforthedetectionofmicrocystin-LRindrinkingwaterwithoutpreconcentration.FoodChemToxicol,40,1677.
10. BarkhudaryanN,GambarovS,GyulbayazyanT,NahapetyanK.(2002)LVV-hemorphin-4modulatesCa2+/calmodulin-dependentpathwaysintheimmunesystembythesamemechanismasinthebrain.JMolNeurosci,18,203.
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。
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1、快讯上是Invotrogen公司的SYRBGOLD,不过500ul/1390元,实在有点贵,为了查用量;
2、结果又查出赛百盛的GoldViewTMDNA染料1ml/100元;
3、还有一种在日光下即可看出DNA条带的上海华舜生物工程有限公司的LightBluedye;
为了一个EB,居然不小心查出三种染料,而对这三种染料的评价也褒贬不一,我个人希望找个毒害小些,而且灵敏度还不能低于EB,价格方面也能让老板接受的来。
而对这三个染料,我有三个问题:
1、SYRBGOLD的用量问题,500ul能用到200次吗(我一般一次用30ml的胶,配在胶中)?
2、GoldViewTMDNA染料的成分是否如有些人所说的就是丫啶橙(能举出确切证据吗)?
3、LightBluedye灵敏度如何?能比得过EB吗?
请各位大侠给予帮助!!
谢谢!!
产品主要应用:点击化学(Clickchemistry)、蛋白质组学研究中的双向荧光差异凝胶电泳(2DDIGE)和实时荧光定量PCR(RealtimePCR)。
菁染料是性能优良的荧光标记染料,摩尔吸光系数在荧光染料中是最高的。N-羟基琥珀酰亚胺酯是最常用的脂肪氨基标记试剂,广泛用于蛋白质、氨基酸多肽、抗体、核酸及其他生物分子的标记和检测。通过改变次甲基链的长度,可改变其荧光发射波长,每增加一个双键,按照Huoffman规则正好红移约100nm。
菁染料Cy3和Cy5已成为基因芯片的首选荧光标记物;另外,Cy5,Cy5.5和Cy7,Cy7.5的吸收在近红外区背景非常低,是荧光强度最高、最稳定的长波长染料,特别适合于活体小动物体内成像。但由于菁染料,尤其是不对称菁染料的合成副反应多,副产物极性相近,产物的分离提纯相当困难。菁染料特别是水溶性菁染料分子极性大,分离提纯越加困难。Lumiprobe供应脂溶性和水溶性菁染料。
相关产品:
产品分子结构可替代染料编号:AGF1371A
6-ROX-N-羟基琥珀酰亚胺酯
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编号:AGF1345A
Cy5.5-N-羟基琥珀酰亚胺酯
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编号:AGF1349A
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Cy7NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103713_8843.jpg[/img]编号:AGF1356A
Cy7.5-N-羟基琥珀酰亚胺酯
Cy7.5NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103756_5406.jpg[/img]编号:AGF1374A
磺酸基-Cy3-N-羟基琥珀酰亚胺酯
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DyLight549
编号:AGF1377A
磺酸基-Cy5-N-羟基琥珀酰亚胺酯
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DyLight649
编号:AGF1379A
磺酸基-Cy7-N-羟基琥珀酰亚胺酯
Sulfo-Cy7NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704104047_6968.jpg[/img]相似系列产品:
羰基活性荧光染料
氨基类染料
巯基反应性染料
羧酸类染料
2、苏丹三 脂肪 橙红
3、苏丹四 脂肪 红
4、双缩脲 蛋白质 紫
5、龙胆紫 染色质 紫
6、碘 淀粉 蓝
7、健那绿 线粒体 绿
8、甲基绿 DNA 绿
9、吡罗红 RNA 红
10、溴麝香草酚蓝 CO2 由蓝变绿再变黄
11、重铬酸钾 酒精 酸性条件下由橙色变成灰绿
12、醋酸洋红(龙胆紫、改良苯酚品红) 染色质 红
13、台盼蓝 检验活死细胞 死细胞会被染成蓝色(不常用)
而且样品中的无水硫酸钠未变色,而做标准曲线的五个和空白对照的变为蓝色了,请高手指教,多谢!
还有,是否变蓝对测定结果有影响吗?
谢了哈
欢迎你!请下次规范发贴:)
健那绿染液是一种活体染液,实验对象必须是活细胞,健那绿可以使活细胞中的线粒体呈现蓝绿色,而细胞质接近无色。
二苯胺
甲基绿派洛宁
还有一种忘了。
假期没法联系老师呵呵。
这三种的作用都是什么。
暂无品牌问答