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AAT Bioquest/Amplite™ Colorimetric Aldehyde Quantitation Kit *Blue Color*/10053/200 Tests
| 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.
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