
Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 403/454 |
MW | N/A |
CAS# | N/A |
Solvent | DMSO |
Storage | F/D/L |
Category | SuperiorLabelingDyes mFluor™DyesandKits |
Related | Generalproteins LabelingviaAminoGroups BiochemicalAssays |
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 |
DeepSequencingAnalysisoftheEha-RegulatedTranscriptomeofEdwardsiellatardaFollowingAcidification
Authors:DGao,NLiu,YLi,YZhang,GLiu
Journal:Metabolomics(LosAngel)(2017):2153--0769
Suramininhibitscullin-RINGE3ubiquitinligases
Authors:KennethWu,RobertAChong,QingYu,JinBai,DonaldESpratt,KevinChing,ChanLee,HaibinMiao,IngerTappin,JerardHurwitz
Journal:ProceedingsoftheNationalAcademyofSciences(2016):E2011--E2018
GlycosaminoglycanmimicrybyCOAMreducesmelanomagrowththroughchemokineinductionandfunction
Authors:HelenePiccard,NeleBerghmans,EvaKorpos,ChrisDillen,IlseVanAelst,SandraLi,ErikMartens,SandraLiekens,SamNoppen,JoVanDamme
Journal:InternationalJournalofCancer(2012):E425--E436
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)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。
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一、探针DNA标记方法
步骤 :
1.10ng~3ugDNA每管,双蒸水定量至总体积15ul
2.沸水水浴10分钟后迅速冰浴
3.加入 5号试剂 2ul , 6号试剂 2ul ,7号试剂 1ul
4.37度1小时~20小时
5.中止反应 加2ul 0.2M EDTA (pH 8.0) 和/或 65度 水浴 10分钟
二、标记效率检测
(一) 试剂配置
Solution Composition Preparation
Washing buffer 0.1 M Maleic acid, 0.15 M NaCl; pH 7.5 (20° C); 0.3% (v/v) Tween 20
Maleic acid buffer 0.1 M Maleic acid, 0.15 M NaCl; adjustwith NaOH (solid) to pH 7.5 (20° C)
Detection buffer 0.1 M Tris-HCl, 0.1 M NaCl, pH 9.5 (20° C)
TE-buffer 10 mM Tris-HCl, 1 mM EDTA, pH 8.0
Blocking stock solution Dissolve Blocking reagent (bottle 10) 10% (w/v) in Maleic
(10 × conc.) acid buffer underconstantly stirring on a heating block(65°C) or heat in a microwave oven,autoclave. The solution remains opaque
Blocking solution Prepare a 1 × working solution by dilutingthe 10 × Blocking solution 1:10in Maleic acid buffer.
Antibody solution Centrifuge Anti-Digoxigenin-AP(vial 8) for 5 min at 10 000 rpm in theoriginal vial prior to each use, and pipet the necessary amount carefully from thesurface. Dilute Anti-
Digoxigenin-AP 1: 5 000 (150 mU/ml) in Blocking solution.
Colorsubstrate Add 40 _l of NBT/BCIP (vial 9) to 2 ml of Detection buffer.
solution Note: Store protected from light!
(二)对照的标记DNA(4号试剂)系列稀释
(三)步骤
1、取以上制备的管2~9各1ul,以及自己标记的探针1ul,点到一小片尼龙膜
2、通过紫外线或者120度半小时使核酸交连到膜上
3、膜置塑料盒中加Maleic acid buffer 20ml ,15~25度轻摇孵育2分钟
4、10ml Blocking solution 孵育30分钟
5、10ml Antibody solution 孵育30分钟
6、10ml Washing buffer 洗2次,每次15分钟
7、10ml Detection buffer中平衡2~5分钟
8、膜在2ml新鲜配制的 Colorsubstrate solution 中避光孵育。显色期间避免摇动
9、中止反应 用TE-buffer或者双蒸水洗5分钟
蛋白酶K预处理
三、样品检测与杂交
(一) 步骤
1、 稀释供试品及阳性对照DNA,点膜
2、 通过紫外线或者120度半小时使核酸交连到膜上
3、 将标记好的探针稀释到约25ng/ml, 煮沸5分钟后迅速冰浴
4、膜放入预热好的预杂交液(3.5ml/100cm2)中,充分混匀,避免起泡
5、倒掉预杂交液,加入杂交液及已变性的探针。杂交温度下至少孵育6小时
四、洗膜
1、2 × SSC, 0.1% SDS , 15-25° C,洗膜2次,每次5分钟。
2、0.5× SSC, 0.1% SDS ,预热至65~68° C,洗膜2次,每次5分钟
五、结果检测
(一) 试剂配制
Solution Composition Preparation
Washing buffer 0.1 M Maleic acid, 0.15 M NaCl; pH 7.5 (20° C); 0.3% (v/v) Tween 20
Maleic acid buffer 0.1 M Maleic acid, 0.15 M NaCl; adjustwith NaOH (solid) to pH 7.5 (20° C)
Detection buffer 0.1 M Tris-HCl, 0.1 M NaCl, pH 9.5 (20° C)
TE-buffer 10 mM Tris-HCl, 1 mM EDTA, pH 8.0
Blocking stock solution Dissolve Blocking reagent (bottle 10) 10% (w/v) in Maleic
(10 × conc.) acid buffer underconstantly stirring on a heating block(65°C) or heat in a microwave oven,autoclave. The solution remains opaque
Blocking solution Prepare a 1 × working solution by dilutingthe 10 × Blocking solution1:10in Maleic acid buffer.
Antibody solution Centrifuge Anti-Digoxigenin-AP(vial 8) for 5 min at 10 000 rpm in theoriginal vial prior to each use, and pipet the necessary amount carefully from thesurface. Dilute Anti-
Digoxigenin-AP 1: 5 000 (150 mU/ml) in Blocking solution.
Colorsubstrate Add 40 _l of NBT/BCIP (vial 9) to 2 ml of Detection buffer.
solution Note: Store protected from light!
(二) 步骤
1、Washing buffer洗膜1~5分钟
2、100ml Blocking solution 孵育30分钟
3、20ml Antibody solution 孵育30分钟
4、100ml Washing buffer 洗2次,每次15分钟
5、20ml Detection buffer中平衡2~5分钟
6、膜在10ml新鲜配制的 Colorsubstrate solution 中避光孵育。显色期间摇动
7、中止反应 用TE-buffer或者双蒸水50ml洗5分钟展开
生物偶联即将两种分子结合在一起而形成一个杂合的偶联物(hybridconjugate),这些偶联结合物(如标记抗体)在世界各大医院和科研机构被广泛使用。一抗标记试剂系列产品极大程度的简化了结合标记流程,是目前世界上操作最简便的抗体标记试剂盒。
·快速:手工操作时间仅30秒·目前世界上操作最简便的抗体标记试剂盒·无需脱盐、透析等步骤·抗体回收率达100%·可提供数十种标记物·可用于WB,IHC,ELISA,免疫荧光、流式细胞术以及荧光共振能量转移检测等·经济实惠,规格多样·可标记ug-mg级抗体量试剂盒组成:
1)1或3(以规格而定)玻管Lightening-Link?Mix混合物。
2)1管LL-X(X为标记物,如PE或HRP等)修饰试剂。
3)1管LL-X萃灭试剂。
操作步骤:
1)在每10ul抗体样品中加入1ul的LL-Modifier修饰试剂,轻轻混匀。
2)打开Ligtening-Link混合物管盖,吸取步骤1已混好的抗体样品至管内冻干混合物中,用吸液器轻轻吸打1-2次,使粉末完全溶解。
3)盖上玻璃瓶盖,室温(20-25°C)避光孵育3小时,或过夜。*
4)孵育3小时后,按每10ul抗体的量加入1ulLL-淬灭试剂,静置30分钟后即可使用。
*不同的标记物,操作步骤略有不同。(操作步骤供参考,以英文原版说明书为准。)
标记结合物的保存:
对于任何新结合物,推荐保持于4°C条件下。如需长期保存,可添加保护剂。
缓冲液及添加剂兼容性:
A.与无胺基缓冲液兼容,包括MES,MOPS,HEPES(浓度10-50mM,pH范围6.5-8.5),LL-Modifier修饰剂提供结合反应的最佳条件。
B.常见的盐(如氯化钠)、螯合剂(EDTA)、糖对偶联反应无影响。
C.叠氮钠(0.02%-0.1%)对结合反应有轻微影响,叠氮钠不可用于HRP标记,其为后者的抑制剂。
D.甘油(upto50%)对结合效率无影响。
E.若无胺基的缓冲液浓度相对较高,pH值在6.6-8.5范围外,则需要加入更多的LL-midifier修饰剂(/10ul抗体),以此来调整pH值。最佳的pH范围为7.3-7.6左右。
F.避免使用含亲核剂的缓冲液,避免还原剂如DTT和2-巯基乙醇,乙醇胺等。
G.浓度达到或稍低于20mM的Tris对结合效率有轻微的影响。
Westernblottingdatacomparingconjugatedandunconjugatedantibodies
货号中文品名英文品名规格AZK0001AColorlockALS磷检测试剂盒PiColorlockALS625/1560assaysAZK0001BColorlockALS磷检测试剂盒PiColorlockALS2500/6250assaysAZK0002AColorlockGold磷检测试剂盒PiColorlockGold625/1560assaysAZK0002BColorlockGold磷检测试剂盒PiColorlockGold2500/6250assaysAZK0003APi结合树脂PiBind5gAZK0004AATP琼脂糖Low(1-2umol/ml)ATPagaroseLow(1-2umol/ml)2mlAZK0004BATP琼脂糖Low(1-2umol/ml)ATPagaroseLow(1-2umol/ml)5mlAZK0005AATP琼脂糖High(>6umol/ml)GTPagaroseHigh(>6umol/ml)2mlAZK0006AATP琼脂糖High(1-2umol/ml)GTPagaroseHigh(>6umol/ml)5mlAZK0007AATP琼脂糖High(8-12umol/ml)ATPagaroseHigh(8-12umol/ml)2mlAZK0008AATP琼脂糖High(8-12umol/ml)ATPagaroseHigh(8-12umol/ml)5mlAZK0009A控制树脂ControlResin2mlAZK0009B控制树脂ControlResin5mlAZK0010AATPase酶检测试剂盒ATPaseassaysystem192assaysAZK0010BATPase酶检测试剂盒ATPaseassaysystem480assaysAZK0010CATPase酶检测试剂盒ATPaseassaysystem5x480assaysAZK0011AGTPase酶检测试剂盒GTPaseassaysystem192assaysAZK0011BGTPase酶检测试剂盒GTPaseassaysystem480assaysAZK0011CGTPase酶检测试剂盒GTPaseassaysystem5x480assaysAZK0013AAP碱性磷酸酶抗体标记试剂盒Lightning-LinkAPconjugationkit3x10μgAPAZK0013BAP碱性磷酸酶抗体标记试剂盒Lightning-LinkAPconjugationkit1x100μgAPAZK0013CAP碱性磷酸酶抗体标记试剂盒Lightning-LinkAPconjugationkit3x100μgAPAZK0013DAP碱性磷酸酶抗体标记试剂盒Lightning-LinkAPconjugationkit1mgAPAZK0014AGOx葡糖氧化酶标记试剂盒Lightning-LinkGOxconjugationkit3x10μgAZK0014BGOx葡糖氧化酶标记试剂盒Lightning-LinkGOxconjugationkit3x100μgAZK0014CGOx葡糖氧化酶标记试剂盒Lightning-LinkGOxconjugationkit1x1mgAZK0015AR-PE(PE)R-藻红蛋白标记试剂盒Lightning-LinkR-Phycoerythrin(R-PE)3x10μgPEAZK0015BR-PE(PE)R-藻红蛋白标记试剂盒Lightning-LinkR-Phycoerythrin(R-PE)1x100μgPEAZK0015CR-PE(PE)R-藻红蛋白标记试剂盒Lightning-LinkR-Phycoerythrin(R-PE)3x100μgPEAZK0015DR-PE(PE)R-藻红蛋白标记试剂盒Lightning-LinkR-Phycoerythrin(R-PE)1mgPEAZK0016AB-PEB-藻红蛋白标记试剂盒Lightning-LinkB-Phycoerythrin(B-PE)1x100μgBPEAZK0016BB-PEB-藻红蛋白标记试剂盒Lightning-LinkB-Phycoerythrin(B-PE)3x100μgBPEAZK0016CB-PEB-藻红蛋白标记试剂盒Lightning-LinkB-Phycoerythrin(B-PE)1mgBPEAZK0017AAPC别藻蓝蛋白标记试剂盒Lightning-LinkAllophycocyanin(APC)3x10μgAPCAZK0017BAPC别藻蓝蛋白标记试剂盒Lightning-LinkAllophycocyanin(APC)1x100μgAPCAZK0017CAPC别藻蓝蛋白标记试剂盒Lightning-LinkAllophycocyanin(APC)3x100μgAPCAZK0017DAPC别藻蓝蛋白标记试剂盒Lightning-LinkAllophycocyanin(APC)1mgAPCAZK0018APerCP标记试剂盒Lightning-LinkPerCP3x10ugAZK0018BPerCP标记试剂盒Lightning-LinkPerCP1x100ugAZK0018CPerCP标记试剂盒Lightning-LinkPerCP3x100ugAZK0018DPerCP标记试剂盒Lightning-LinkPerCP1x1mgAZK0019A荧光素标记试剂盒Lightning-LinkFluorescein3reactions(eachupto20ug)AZK0019B荧光素标记试剂盒Lightning-LinkFluorescein3reactions(eachupto200ug)AZK0019C荧光素标记试剂盒Lightning-LinkFluorescein1reaction(upto2mg)AZK0020A罗丹明标记试剂盒Lightning-LinkRhodamine3reactions(eachupto20ug)AZK0020B罗丹明标记试剂盒Lightning-LinkRhodamine3reactions(eachupto200ug)AZK0020C罗丹明标记试剂盒Lightning-LinkRhodamine1reaction(upto2mg)AZK0021AAMCA标记试剂盒Lightning-LinkAMCA3reactions(eachupto20ug)AZK0021BAMCA标记试剂盒Lightning-LinkAMCA3reactions(eachupto200ug)AZK0021CAMCA标记试剂盒Lightning-LinkAMCA1reaction(upto2mg)AZK0022ATexasRed标记试剂盒Lightning-LinkTexasRed3reactions(eachupto20ug)AZK0022BTexasRed标记试剂盒Lightning-LinkTexasRed3reactions(eachupto200ug)AZK0022CTexasRed标记试剂盒Lightning-LinkTexasRed1reaction(upto1mgscale)AZK0023ACy3染料标记试剂盒Lightning-LinkCy33reactions(eachupto20ug)AZK0023BCy3染料标记试剂盒Lightning-LinkCy33reactions(eachupto200ug)AZK0023CCy3染料标记试剂盒Lightning-LinkCy31reaction(upto2mg)AZK0024ACy5染料标记试剂盒Lightning-LinkCy53reactions(eachupto20ug)AZK0024BCy5染料标记试剂盒Lightning-LinkCy53reactions(eachupto200ug)AZK0024CCy5染料标记试剂盒Lightning-LinkCy51reaction(upto2mg)AZK0025ACy5.5染料标记试剂盒Lightning-LinkCy5.53reactions(eachupto200ug)AZK0025BCy5.5染料标记试剂盒Lightning-LinkCy5.51reaction(upto2mg)AZK0026ACy7染料标记试剂盒Lightning-LinkCy73reactions(eachupto20ug)AZK0026BCy7染料标记试剂盒Lightning-LinkCy73reactions(eachupto200ug)AZK0026CCy7染料标记试剂盒Lightning-LinkCy71reaction(upto2mg)AZK0027ACy3.5染料标记试剂盒Lightning-LinkCy3.53reactions(eachupto200ug)AZK0027BCy3.5染料标记试剂盒Lightning-LinkCy3.51reaction(upto2mg)AZK0028AAtto390荧光染料标记试剂盒Lightning-Linkatto3903reactions(eachupto20ug)AZK0028BAtto390荧光染料标记试剂盒Lightning-Linkatto3903reactions(eachupto200ug)AZK0028CAtto390荧光染料标记试剂盒Lightning-Linkatto3901reaction(upto2mg)AZK0029AAtto425荧光染料标记试剂盒Lightning-Linkatto4253reactions(eachupto20ug)AZK0029BAtto425荧光染料标记试剂盒Lightning-Linkatto4253reactions(eachupto200ug)AZK0029CAtto425荧光染料标记试剂盒Lightning-Linkatto4251reaction(upto2mg)AZK0030AAtto465荧光染料标记试剂盒Lightning-Linkatto4653reactions(eachupto20ug)AZK0030BAtto465荧光染料标记试剂盒Lightning-Linkatto4653reactions(eachupto200ug)AZK0030CAtto465荧光染料标记试剂盒Lightning-Linkatto4651reaction(upto2mg)AZK0031AAtto488荧光染料标记试剂盒Lightning-Linkatto4883reactions(eachupto20ug)AZK0031BAtto488荧光染料标记试剂盒Lightning-Linkatto4883reactions(eachupto200ug)AZK0031CAtto488荧光染料标记试剂盒Lightning-Linkatto4881reaction(upto2mg)AZK0032AAtto532荧光染料标记试剂盒Lightning-Linkatto5323reactions(eachupto20ug)AZK0032BAtto532荧光染料标记试剂盒Lightning-Linkatto5323reactions(eachupto200ug)AZK0032CAtto532荧光染料标记试剂盒Lightning-Linkatto5321reaction(upto2mg)AZK0033AAtto565荧光染料标记试剂盒Lightning-Linkatto5653reactions(eachupto20ug)AZK0033BAtto565荧光染料标记试剂盒Lightning-Linkatto5653reactions(eachupto200ug)AZK0033CAtto565荧光染料标记试剂盒Lightning-Linkatto5651reaction(upto2mg)AZK0034AAtto590荧光染料标记试剂盒Lightning-Linkatto5903reactions(eachupto20ug)AZK0034BAtto590荧光染料标记试剂盒Lightning-Linkatto5903reactions(eachupto200ug)AZK0034CAtto590荧光染料标记试剂盒Lightning-Linkatto5901reaction(upto2mg)AZK0035AAtto594荧光染料标记试剂盒Lightning-Linkatto5943reactions(eachupto20ug)AZK0035BAtto594荧光染料标记试剂盒Lightning-Linkatto5943reactions(eachupto200ug)AZK0035CAtto594荧光染料标记试剂盒Lightning-Linkatto5941reaction(upto2mg)AZK0036AAtto633荧光染料标记试剂盒Lightning-Linkatto6333reactions(eachupto20ug)AZK0036BAtto633荧光染料标记试剂盒Lightning-Linkatto6333reactions(eachupto200ug)AZK0036CAtto633荧光染料标记试剂盒Lightning-Linkatto6331reaction(upto2mg)AZK0037AAtto637荧光染料标记试剂盒Lightning-Linkatto6373reactions(eachupto200ug)AZK0037BAtto637荧光染料标记试剂盒Lightning-Linkatto6371reaction(upto2)mgAZK0038AAtto655荧光染料标记试剂盒Lightning-Linkatto6553reactions(eachupto20ug)AZK0038BAtto655荧光染料标记试剂盒Lightning-Linkatto6553reactions(eachupto200ug)AZK0038CAtto655荧光染料标记试剂盒Lightning-Linkatto6551reaction(upto2mg)AZK0039AAtto680荧光染料标记试剂盒Lightning-Linkatto6803reactions(eachupto20ug)AZK0039BAtto680荧光染料标记试剂盒Lightning-Linkatto6803reactions(eachupto200ug)AZK0039CAtto680荧光染料标记试剂盒Lightning-Linkatto6801reaction(upto2mg)AZK0040AAtto700荧光染料标记试剂盒Lightning-Linkatto7003reactions(eachupto20ug)AZK0040BAtto700荧光染料标记试剂盒Lightning-Linkatto7003reactions(eachupto200ug)AZK0040CAtto700荧光染料标记试剂盒Lightning-Linkatto7001reaction(upto2mg)AZK0041AFluoProbesDyes547H荧光探针染料标记试剂盒Lightning-LinkFluoProbes547H3reactions(eachupto20ug)AZK0041BFluoProbesDyes547H荧光探针染料标记试剂盒Lightning-LinkFluoProbes547H3reactions(eachupto200ug)AZK0041CFluoProbesDyes547H荧光探针染料标记试剂盒Lightning-LinkFluoProbes547H1reaction(upto2mg)AZK0042AFluoProbesDyes594荧光探针染料标记试剂盒Lightning-LinkFluoProbes5943reactions(eachupto20ug)AZK0042BFluoProbesDyes594荧光探针染料标记试剂盒Lightning-LinkFluoProbes5943reactions(eachupto200ug)AZK0042CFluoProbesDyes594荧光探针染料标记试剂盒Lightning-LinkFluoProbes5941reaction(upto2mg)AZK0043AFluoProbesDyes647H荧光探针染料标记试剂盒Lightning-LinkFluoProbes647H3reactions(eachupto20ug)AZK0043BFluoProbesDyes647H荧光探针染料标记试剂盒Lightning-LinkFluoProbes647H3reactions(eachupto200ug)AZK0043CFluoProbesDyes647H荧光探针染料标记试剂盒Lightning-LinkFluoProbes647H1reaction(upto2mg)AZK0044AFluoProbesDyes682荧光探针染料标记试剂盒Lightning-LinkFluoProbes6823reactions(eachupto20ug)AZK0044BFluoProbesDyes682荧光探针染料标记试剂盒Lightning-LinkFluoProbes6823reactions(eachupto200ug)AZK0044CFluoProbesDyes682荧光探针染料标记试剂盒Lightning-LinkFluoProbes6821reaction(upto2mg)AZK0045AFluoProbesDyes752荧光探针染料标记试剂盒Lightning-LinkFluoProbes7523reactions(eachupto20ug)AZK0045BFluoProbesDyes752荧光探针染料标记试剂盒Lightning-LinkFluoProbes7523reactions(eachupto200ug)AZK0045CFluoProbesDyes752荧光探针染料标记试剂盒Lightning-LinkFluoProbes7521reaction(upto2mg)AZK0046APE/Cy5串联染料Lightning-LinkPE/Cy53x10ugAZK0046BPE/Cy5串联染料Lightning-LinkPE/Cy51x100ugAZK0046CPE/Cy5串联染料Lightning-LinkPE/Cy53x100ugAZK0046DPE/Cy5串联染料Lightning-LinkPE/Cy51x1mgAZK0047APE/Cy5.5串联染料Lightning-LinkPE/Cy5.53x10ugAZK0047BPE/Cy5.5串联染料Lightning-LinkPE/Cy5.51x100ugAZK0047CPE/Cy5.5串联染料Lightning-LinkPE/Cy5.53x100ugAZK0047DPE/Cy5.5串联染料Lightning-LinkPE/Cy5.51x1mgAZK0048APE/Cy7串联染料Lightning-LinkPE/Cy73x10ugAZK0048BPE/Cy7串联染料Lightning-LinkPE/Cy71x100ugAZK0048CPE/Cy7串联染料Lightning-LinkPE/Cy73x100ugAZK0048DPE/Cy7串联染料Lightning-LinkPE/Cy71x1mgAZK0049APE/Cy5.5串联染料Lightning-LinkPerCP/Cy5.53x10ugAZK0049BPE/Cy5.5串联染料Lightning-LinkPerCP/Cy5.51x100ugAZK0050APE/TexasRed串联染料Lightning-LinkPE/TexasRed3x10ugAZK0050BPE/TexasRed串联染料Lightning-LinkPE/TexasRed1x100ugAZK0050CPE/TexasRed串联染料Lightning-LinkPE/TexasRed3x100ugAZK0050DPE/TexasRed串联染料Lightning-LinkPE/TexasRed1x1mgAZK0051APE/Atto594串联染料Lightning-Link?PE/Atto5943x10ugAZK0051BPE/Atto594串联染料Lightning-Link?PE/Atto5941x100ugAZK0051CPE/Atto594串联染料Lightning-Link?PE/Atto5943x100ugAZK0051DPE/Atto594串联染料Lightning-Link?PE/Atto5941x1mgAZK0052APerCP/Cy5.5串联染料Lightning-LinkPerCP/Cy5.53x100ugAZK0052BPerCP/Cy5.5串联染料Lightning-LinkPerCP/Cy5.51x1mgAZK0052CPerCP/Cy5.5串联染料Lightning-LinkAPC/Cy5.53x10ugAZK0052DPerCP/Cy5.5串联染料Lightning-LinkAPC/Cy5.51x100ugAZK0052EPerCP/Cy5.5串联染料Lightning-LinkAPC/Cy5.53x100ugAZK0052FPerCP/Cy5.5串联染料Lightning-LinkAPC/Cy5.51x1mgAZK0053ALightning-LinkAPC/Cy7串联染料Lightning-LinkAPC/Cy73x10ugAZK0053BLightning-LinkAPC/Cy7串联染料Lightning-LinkAPC/Cy71x100ugAZK0053CLightning-LinkAPC/Cy7串联染料Lightning-LinkAPC/Cy73x100ugAZK0053DLightning-LinkAPC/Cy7串联染料Lightning-LinkAPC/Cy71x1mgAZK0054A生物素化结合试剂盒typeALightning-LinkBiotinconjugationkittypeA3reactions(upto20μgAb)AZK0054B生物素化结合试剂盒typeALightning-LinkBiotinconjugationkittypeA3reactions(upto200μgAb)AZK0054C生物素化结合试剂盒typeALightning-LinkBiotinconjugationkittypeA1reaction(upto2mgAb)AZK0055A生物素化结合试剂盒typeBLightning-LinkBiotinconjugationkittypeB3reactions(upto20μgAb)AZK0055B生物素化结合试剂盒typeBLightning-LinkBiotinconjugationkittypeB3reactions(upto200μgAb)AZK0055C生物素化结合试剂盒typeBLightning-LinkBiotinconjugationkittypeB1reaction(upto2mgAb)AZK0056A链亲和素标记试剂盒Lightning-LinkStreptavidinconjugationkit3x10ugStreptavidinAZK0056B链亲和素标记试剂盒Lightning-LinkStreptavidinconjugationkit3x100ugStreptavidinAZK0056C链亲和素标记试剂盒Lightning-LinkStreptavidinconjugationkit1x1mgStreptavidinAZK0057A亲和素标记试剂盒Lightning-LinkAvidinconjugationkit3x10ugAvidinAZK0057B亲和素标记试剂盒Lightning-LinkAvidinconjugationkit3x100ugAvidinAZK0057C亲和素标记试剂盒Lightning-LinkAvidinconjugationkit1x1mgAvidinAZK0058A快速荧光素标记Lightning-Link?RapidFluorescein3xAblabelings(each10-20ugscale)AZK0058B快速荧光素标记Lightning-Link?RapidFluorescein3xAblabelings(each100-200ugscale)AZK0058C快速荧光素标记Lightning-Link?RapidFluorescein1xAblabeling(1scale)AZK0059A快速Cy3标记Lightning-Link?RapidCy33xAblabelings(each10-20ugscale)AZK0059B快速Cy3标记Lightning-Link?RapidCy33xAblabelings(each100-200ugscale)AZK0059C快速Cy3标记Lightning-Link?RapidCy31xAblabeling(1scale)AZK0060A快速Cy5标记Lightning-Link?RapidCy53xAblabelings(each10-20ugscale)AZK0060B快速Cy5标记Lightning-Link?RapidCy53xAblabelings(each100-200ugscale)AZK0060C快速Cy5标记Lightning-Link?RapidCy51xAblabeling(1scale)AZK0061A快速Atto488标记Lightning-Link?RapidAtto4883xAblabelings(each10-20ugscale)AZK0061B快速Atto488标记Lightning-Link?RapidAtto4883xAblabelings(each100-200ugscale)AZK0061C快速Atto488标记Lightning-Link?RapidAtto4881xAblabeling(1scale)AZK0062A快速Atto633标记Lightning-Link?RapidAtto6333xAblabelings(each10-20ugscale)AZK0062B快速Atto633标记Lightning-Link?RapidAtto6333xAblabelings(each100-200ugscale)AZK0062C快速Atto633标记Lightning-Link?RapidAtto6331xAblabeling(1scale)AZK0063A快速Atto565标记Lightning-Link?RapidAtto5653xAblabelings(each10-20ugscale)AZK0063B快速Atto565标记Lightning-Link?RapidAtto5653xAblabelings(each100-200ugscale)AZK0063C快速Atto565标记Lightning-Link?RapidAtto5651xAblabeling(1scale)AZK0064A快速Atto594标记Lightning-Link?RapidAtto5943xAblabelings(each10-20ugscale)AZK0064B快速Atto594标记Lightning-Link?RapidAtto5943xAblabelings(each100-200ugscale)AZK0064C快速Atto594标记Lightning-Link?RapidAtto5941xAblabeling(1scale)AZK0065A快速罗丹明标记Lightning-Link?RapidRhodamine3xAblabelings(each10-20ugscale)AZK0065B快速罗丹明标记Lightning-Link?RapidRhodamine3xAblabelings(each100-200ugscale)AZK0065C快速罗丹明标记Lightning-Link?RapidRhodamine1xAblabeling(1scale)AZK0066A快速Atto390标记Lightning-Link?RapidAtto3903xAblabelings(each10-20ugscale)AZK0066B快速Atto390标记Lightning-Link?RapidAtto3903xAblabelings(each100-200ugscale)AZK0066C快速Atto390标记Lightning-Link?RapidAtto3901xAblabeling(1scale)AZK0067A快速DyLight350标记Lightning-Link?RapidDyLight3503xAblabelings(each10-20ugscale)AZK0068A快速DyLight350素标记Lightning-Link?RapidDyLight3503xAblabelings(each100-200ugscale)AZK0069A快速DyLight350标记Lightning-Link?RapidDyLight3501xAblabeling(1scale)AZK0070A快速DyLight405标记Lightning-Link?RapidDyLight4053xAblabelings(each10-20ugscale)AZK0070B快速DyLight405标记Lightning-Link?RapidDyLight4053xAblabelings(each100-200ugscale)AZK0070C快速DyLight405标记Lightning-Link?RapidDyLight405
探针
ddH2O36.25ul
buffer(瓶3)5ul
核苷酸混合液(含DIG-dUTP)(瓶2)2.5ul
dNTPStockSolution(瓶4)2.5ul
引物F1ul(10pmol/ul)
R1ul(10pmol/ul)
酶(瓶1)0.75ul
DNA1ul(100pg)
程序:35个循环,退火温度55度。
第一次通过探针标记得到了相应的PCR标记产物,亮度和对照相当,片段大小比标记的探针模板大,证明整个过程应该没有问题。
但一个月后重新用相同的体系,相同的程序,只是探针模板的量不同却什么都扩不出来。不知道问题出在哪了!

