Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 620/None |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | SmallMoleculeDetection DiagnosticMolecules |
Related | RedoxEnzymes BiochemicalAssays |
Note:Thawallthekitcomponentstoroomtemperaturebeforestartingtheexperiment.
1.Prepare2XAldeView™Bluereactionmixture:
Add5mLofAssayBuffer(ComponentB)intoonebottleofAldeView™Blue(ComponentA)tomakeAldeView™Bluereactionmixture.
Note:5mLof2XAldeView™Bluereactionmixtureisenoughforoneplate.Thereactionmixtureisnotstable,andbestusedwithin2hours.
2.Prepareserialdilutionsofaldehydestandard(0to100µM):
2.1 Add1mLofAssayBuffer(ComponentB)intothevialofAldehydeStandard(ComponentD)tomakea10mMaldehydestandardstocksolution.
Note:Theunused10mMaldehydestandardstocksolutionshouldbedividedintosingleusealiquotsandstoredat
-20oC.
2.2 Take100μLof10mMaldehydestandardstocksolution(fromStep2.1)toperform1:100,and1:2serialdilutionstoget100,50,25,12.5,6.25,3.125,1.56and0μMserialdilutionswithAssayBuffer(ComponentB).
2.3 Addseriallydilutedaldehydestandardsandaldehyde-containingtestsamplesintoawhite96-wellmicroplatewithclearbottomasdescribedinTables1and2.
Table1.LayoutofAldehydeStandardsandtestsamplesinawhite96-wellmicroplatewithclearbottom
BL | BL | TS | TS | …. | …. |
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AS1 | AS1 | …. | …. | …. | …. |
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AS2 | AS2 |
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AS3 | AS3 |
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AS4 | AS4 |
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AS5 | AS5 |
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AS6 | AS6 |
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AS7 | AS7 |
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Note:AS=AldehydeStandards,BL=BlankControl,TS=TestSamples.
Table2.Reagentcompositionforeachwell
AldehydeStandards | BlankControl | TestSample |
Serialdilutions*:50μL | AssayBuffer:50μL | 50μL |
*Note:Addtheseriallydilutedaldehydestandardsfrom1.56μMto100μMintowellsfromAS1toAS7induplicate.
3.Runaldehydeassay:
3.1 Add50μLof 2XAldeView™Bluereactionmixture(fromStep1)intoeachwellofaldehydestandard,blankcontrol,andtestsamples(seeStep2.3)tomakethetotalaldehydeassayvolumeof100µL/well.
Note:Fora384-wellplate,add12.5μLoftestsampleand12.5μLof2XAldeView™Bluereactionmixtureintoeachwell.
3.2 Incubatethereactionmixtureatroomtemperaturefor20-30minutes(protectedfromlight).
3.3 Add50μLofAldeView™BlueEnhancer(ComponentC)intoeachwell.
Note:Fora384-wellplate,add25μLofAldeView™BlueEnhancerintoeachwell.
3.4 Monitortheabsorbanceincreaseataround620to660nm(Maxat620nm)usinganabsorbanceplatereader.
References&Citations | CitationExplorer |
BiologicalActivityofPeptide-conjugatedPolyionComplexMatricesConsistingofAlginateandChitosan
Authors:ChikaraFujimori,JunKumai,KyotaroNakamura,YingziGu,FumihikoKatagiri,KentaroHozumi,YamatoKikkawa,MotoyoshiNomizu
Journal:PeptideScience(2016)
HepaticDeficiencyofAugmenterofLiverRegenerationExacerbatesAlcohol-InducedLiverInjuryandPromotesFibrosisinMice
Authors:SudhirKumar,JiangWang,RichaRani,ChandrashekharRGandhi
Journal:PloSone(2016):e0147864
Integratedself-assemblingdrugdeliverysystempossessingdualresponsiveandactivetargetingfororthotopicovariancancertheranostics
Authors:Chun-JuiLin,Chen-HsiangKuan,Li-WenWang,Hsi-ChinWu,YunchingChen,Chien-WenChang,Rih-YangHuang,Tzu-WeiWang
Journal:Biomaterials(2016):12--26
Fiber-opticproteasesensorbasedonthedegradationofthingelatinfilms
Authors:BastienSchyrr,StéphanieBoder-Pasche,RéalIscher,RitaSmajda,GuyVoirin
Journal:SensingandBio-SensingResearch(2015):65--73
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临床实验室对商品定量试剂盒分析性能的验证.pdf(17431.6k)
1.模板提取(一般为RNA):Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定:分光光度计或NanoDrop
3.逆转录:逆转录试剂盒(或者一步法试剂盒),这一步可以用普通PCR做,也可以用水域做。
4.荧光定量PCR试剂:通常有用SYBR Green Mix做的,但是这里建议你用EvaGreen做,灵敏度和平行性都要好于SYBR Green,并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他:除了以上的那些还需要离心管、PCR管或板(Axygen反应比较好)、移液枪等,暂时就想到这么多。
至少可以说同一种蛋白定量试剂盒并不适用于所有细胞
蛋白定量试剂盒有分Bradford,BCA,Lowry等,每一种对不同的细胞都有独特的适用范围和灵敏度。所以需要多种蛋白定量试剂盒对比检测才能让结果准确
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