>
产品中心 >
Other_kits >
AAT Bioquest/PhosphoWorks™ Colorimetric Phosphate Assay Kit *Blue Color*/21665/1000 Tests
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 650/None |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | SmallMoleculeDetection Anions |
| Related | ATPases Phosphoproteins BiochemicalAssays |
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.RunPhosphoWorks™colorimetricphosphateassay:
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,eithermultiplethevolumeofsampleandMGPlus™Reagent(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
