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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/PhosphoWorks™ Colorimetric Phosphate Assay Kit *Blue Color*/21665/1000 Tests
产品编号:21665
市  场 价:¥56560.00
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:$2828.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/PhosphoWorks™ Colorimetric Phosphate Assay Kit *Blue Color*/21665/1000 Tests
商品介绍
Overview
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Ex/Em(nm)650/None
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategorySmallMoleculeDetection
Anions
RelatedATPases
Phosphoproteins
BiochemicalAssays
PhosphateisinvolvedinmanyBIOLOGicalreactions.Forexample,phosphatases,ATPasesandseveralotherenzymescatalyzereactionsinwhichinorganicphosphate(Pi)isreleasedfromasubstrate.ThisPhosphoWorks™PhosphateAssayKithasbeendevelopedformeasuringtheactivityofanyPi-generatingenzyme.ThekitisformulatedtogivesensitivedetectionofPi,providinganalternativetohazardousrADIoactivemethodsandotherlesssensitivecolorimetricassays.ThemeasurementofPiisbasedonthechangeinabsorbanceofamalachitegreenderivativeinthepresenceofmolyBDate.Unlikeothermalachitedyeformulations,thiskitgivesacompletelystableend-pointsignalthatisnotpronetoprecipitation.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Prepareassayreagents:

Caution:Phosphate-containingbuffersshouldbeavoidedwhenpreparingthesamples!

1.1   Warmallcomponentsatroomtemperaturebeforeuse.

 

1.2   Itisstronglyrecommendthatclearmicroplatesorcuvettesbeusedtoachievethebestresults.

 

2.Prepareseriallydilutedphosphatestandardsand/ortestsamples:

2.1   Add50μLof1mMphosphatestandard(ComponentA)in950μLofdeionizedwaterorenzymereactionbuffertogeta50μMphosphatestandardsolution.

 

2.2   Take200μLof50μMphosphatestandardsolutiontoperform1:2serialdilutionstoget25,12.5,6.25,3.125,1.56,and0.78μMseriallydilutedphosphatestandards.

 

2.3   Addphosphate-containingtestsamplesintoaclear96-wellmicroplateasdescribedinTables1and2.

 

Table1Layoutofphosphatestandardsandtestsamplesinaclear96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

PS1

PS1

….

….

….

….

 

 

 

 

 

 

PS2

PS2

 

 

 

 

 

 

 

 

 

 

PS3

PS3

 

 

 

 

 

 

 

 

 

 

PS4

PS4

 

 

 

 

 

 

 

 

 

 

PS5

PS5

 

 

 

 

 

 

 

 

 

 

PS6

PS6

 

 

 

 

 

 

 

 

 

 

PS7

PS7

 

 

 

 

 

 

 

 

 

 

Note:PS=PhosphateStandards,BL=BlankControl,TS=TestSamples.

 

Table2.Reagentcompositionforeachwell

PhosphateStandard

BlankControl

TestSample

SerialDilutions*:80μL

Phosphate-freewaterorbuffer:80μL

80μL

Note:*Addtheserialdilutionsofphosphatestandardfrom0.1μMto50μMintowellsfromPS1toPS7induplicate.

 

3.RunPhosphoWorkscolorimetricphosphateassay:

3.1   ShakeMGPlus™Reagent(ComponentB)wellbeforeuse.Add20μL/wellofMGPlus™Reagent(ComponentB)intothewellsofphosphatestandards,blankcontrol,andtestsamples.Mixthereagentsthoroughly.

Note:Fora384-wellplate,add40μLofsampleand10μLofMGPlus™Reagent(ComponentB)intoeachwell.

 

3.2   Ablue-greencolorwilldevelopinthephosphate-containingwellsin10to40minutes.Monitorabsorbanceat600-660nmwithanabsorbancemicroplatereaderoraspectrophotometer.

Note1:Athighphosphateconcentration(>100μM),precipitatesmayform.Diluteyoursamplesandredotheassays.

Note2:Forcuvetteassaythatrequiresthetotalvolumelargerthan100μL,eithermultiplethevolumeofsampleandMGPlusReagent(ComponentB)proportionallyordilutethefinalreactionmixturewith1 MH2OS4 or1MHClbeforemeasuringtheabsorption.

References&Citations
CitationExplorer

VariationofboneformingABIlitywiththephysicochemicalpropertiesofcalciumphosphatebonesubstitutes
Authors:RongquanDuan,DavideBarbieri,XiaomanLuo,JieWeng,ChongyunBao,JoostDeBruijn,HuipinYuan
Journal:BiomaterialsScience(2017)

Influenceofsurfacemicrostructureandchemistryonosteoinductionandosteoclastogenesisbybiphasiccalciumphosphatediscs.
Authors:NLDavison,JSu,HYuan,JJJPvandenBeucken,JDdeBruijn
Journal:(2015)

Submicron-scalesurfacearchitectureoftricalciumphosphatedirectsosteogenesisinvitroandinvivo
Authors:NLDavison,XLuo,TSchoenmaker,VEverts,HYuan,FBarrere-deGroot,JDdeBruijn
Journal:EurCellMater(2014)

Zincincalciumphosphatemediatesboneinduction:invitroandinvivomodel
Authors:XiaomanLuo,DavideBarbieri,NoelDavison,YonggangYan,JoostDdeBruijn,HuipinYuan
Journal:Actabiomaterialia(2014):477--485


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