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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/MESG *Phosphate assay reagent*/21600/5 mg
产品编号:21600
市  场 价:¥21760.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$1088.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/MESG *Phosphate assay reagent*/21600/5 mg
商品介绍
Overview
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Ex/Em(nm)330/None
MW313.34
CAS#N/A
SolventDMSO
StorageF/D/L
CategorySmallMoleculeDetection
Anions
RelatedCellSignalingMolecules
BiochemicalAssays
InthepresenceofinorganicphosphateMESGisconvertedto2-amino-6-mercapto-7-methlpurinebypurinenucleosidephosphorylase(EC2.4.2.1)withabsorptionwavelengthshifttored.Thisfeaturehasbeenusedtoquantifyingphosphatespectrophotometrically.WeofferaconvenientMESG-basedphosphateassaykit(#21659).

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Prepareassayreagents:

1.1   Thawallthefourcomponentsatroomtemperaturebeforeuse.

1.2   PrepareMESGSubstrate(ComponentB)Solution:Add500μLofddH2OtothevialofMESGSubstrate(ComponentB).MixwellbyvortexingtogetMESGSubstrateSolution.

Note:250ulisenoughforoneplatemakesingleusedaliquosandstoreitat-20°CImmediately.,

1.3   PreparePurineNucleosidePhosphorylase(ComponentC)Solution:Add100μLofddH2OtothevialofPurineNucleosidePhosphorylase(PNP;ComponentC).MixwellbyvortexingtogetPurineNucleosidePhosphorylaseSolution.

1.4   PrepareAssaySolution:AddthewholevolumeofMESGSubstrateSolution(fromStep1.2)andPurineNucleosidePhosphorylaseSolution(fromStep1.3)intothebottleofAssayBuffer(ComponentA),mixwelltogettheassaysolution.Placetheassaysolutiononice.

Note1:ThisAssaySolutionisstableforatleast4hoursonice.Itisnotrecommendedtofreezetheassaysolutionforanotherassay.

Note2:Toachievethedesirableresults,UV-transparentplatesorcuvettesarerequired.

Note3:DuetothehighsensitivityofthisassaytoPi,itisextremelyimportanttousePi-freelaboratorywareand

 

2.Prepareseriallydilutedphosphatestandardsand/ortestsamples:

2.1   PreparePhosphateStandard:Add50μLof1mMKH2PO4(ComponentD)into950μLofdeionizedwaterorenzymereactionbuffertogive50μMphosphatestandardsolution.

2.2   Take200μLof50μMphosphatestandardsolutiontoperform1:2serialdilutionstogive25,12.5,6.25,3.125,1.56,and0.78μMseriallydilutedphosphatestandards.

 

2.3   Addphosphate-containingtestsamplesand/orphosphatestandardsintoaclearUV-transparent96-wellmicroplateaccordingtoTables1and2.

 

Table1LayoutofphosphatestandardsandtestsamplesinaclearUV-transparent96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

PS1

PS1

….

….

….

….

 

 

 

 

 

 

PS2

PS2

 

 

 

 

 

 

 

 

 

 

PS3

PS3

 

 

 

 

 

 

 

 

 

 

PS4

PS4

 

 

 

 

 

 

 

 

 

 

PS5

PS5

 

 

 

 

 

 

 

 

 

 

PS6

PS6

 

 

 

 

 

 

 

 

 

 

PS7

PS7

 

 

 

 

 

 

 

 

 

 

Note:PS=PhosphateStandard,BL=BlankControl,TS=TestSample.

Table2Reagentcompositionforeachwell

PhosphateStandard

BlankControl

TestSample

SerialDilutions*:50μL

Phosphate-freewaterorbuffer:50μL

50μL

*Note:Addtheserialdilutionsofphosphatestandardfrom0.1μMto50μMintowellsfromPS1toPS7.

3.RunPhosphoWorksMESGphosphateassay:

3.1   Add50μL/wellofAssaySolution(fromStep1.4)intothewellsofphosphatestandards,blankcontrol,andtestsamples.Mixthereagentsthoroughly.

Note:Fora384-wellplate,add25μLofsampleand25μLofAssaySolutionintoeachwell.

3.2   Incubateatroomtemperaturefor30minutes.Monitortheabsorbancewithamicroplatereaderorspectrophotometerat360nm.

Note:Forcuvetteassaythatrequiresthetotalvolumelargerthan100μL,multiplethevolumeofsampleandassayreagentproportionallybeforemeasuringtheabsorption.

References&Citations
CitationExplorer

Sacrificialcrystaltemplatingofhyaluronicacid-basedhydrogels
Authors:RichelleCThomas,PaulEChung,ShanPModi,JohnGHardy,ChristineESchmidt
Journal:EuropeanPolymerJournal(2016)

Osteogeniccellculturescannotutilizeexogenoussourcesofsyntheticpolyphosphateformineralization
Authors:MarianneBAriganello,SidneyOmelon,FABIoVariola,RimaMWazen,PierreMoffatt,AntonioNanci
Journal:Journalofcellularbiochemistry(2014):2089--2102


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