Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 571/585 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | SmallMoleculeDetection Anions |
Related | ATPases Phosphoproteins |
Spectrum | AdvancedSpectrumViewer |
Note:Toachievethebestresults,it’sstronglyrecommendedtousetheblackplates.
1.PreparePhosphateSensorworkingsolution:
1.1 Thawoneofeachkitcomponentatroomtemperaturebeforeuse.
1.2 Make125XPhosphateSensorstocksolution:Add20μLofDMSOintoPhosphateSensor(ComponentB)tomake125XPhosphateSensorstocksolution.
1.3 MakePhosphateSensorworkingsolution:Transfer20μLof125XPhosphateSensorstocksolution(fromStep1.2)into2.5mLofsterileH2O,andmixwell
Note1:AvoiddirectexposureofPhosphateSensor(ComponentB)tolight.
Note2:AliquotandstoretheunusedAssayBuffer(ComponentA)andPhosphateSensorstocksolution(fromStep1.2)at-20oC.Avoidrepeatedfreeze/thawcyclesandpotentialPicontamination.
Note3:DuetothehighsensitivityofthisassaytoPi,itisextremelyimportanttousePi-freelaboratorywareandreagents.
2.Prepareseriallydilutedphosphatestandardsandtestsamples:
2.1 Preparephosphatestandard:Add50μLof1mMKH2PO4(ComponentC)into950μLofdeionizedwaterorenzymereactionbuffertogeta50μMphosphatestandardsolution.Thentake200μLof50μMphosphatestandardsolutiontoperform1:2serialdilutionstoget25,12.5,6.25,3.125,1.56,and0.78μMseriallydilutedphosphatestandards.
2.2 Addphosphatecontainingsamplesandseriallydilutedphosphatestandardsintoasolidblack96-wellmicroplateaccordingtoTables1and2.
Table1Layoutofphosphatestandardsandtestsamplesinasolidblack96-wellmicroplate
BL | BL | TS | TS | …. | …. |
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PS1 | PS1 | …. | …. | …. | …. |
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PS2 | PS2 |
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PS3 | PS3 |
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PS4 | PS4 |
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PS5 | PS5 |
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PS6 | PS6 |
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PS7 | PS7 |
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Note:PS=PhosphateStandard,BL=BlankControl,TS=TestSample.
Table2Reagentcompositionforeachwell
PhosphateStandards | BlankControl | TestSample |
SerialDilutions*:40μL | H2OorBuffer:40μL | 40μL |
Note:*Addtheserialdilutionsofphosphatestandardfrom0.1μMto50μMintowellsfromPS1toPS7.
3.RunPhosphoWorksTMfluorimetricphosphateassay:
Warning:RunthephosphateassayatpH6.5to7.4.
3.1 Add40μLofAssayBuffer(ComponentA)and20µLofPhosphateSensorworkingsolution(fromStep1.3)intoeachwellofphosphatestandard,blankcontrol,andtestsamples(seeStep2.2)tomakethetotalphosphateassayvolumeof100µL/well.
Note:Fora384-wellplate,add20μLofsample,20μLofAssayBuffer(ComponentA)and10µLofPhosphateSensorworkingsolution(fromStep1.3)intoeachwell.
3.2 Incubatethereactionmixtureatroomtemperaturefor15minutesto1hour.
3.3 MonitorthefluorescenceincreasewithafluorescenceplatereaderatEx/Em=540/590nm.
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
AAT Bioquest常用产品报价单
产品名 | 货号 | 公司分类 | 商城分类 | 美金价 |
AAT Bioquest/Fluorescein, disodium salt *CAS 518-47-8*/2/100 mg | 2 | iFluor Rapid Tests | 酸碱缓冲液 | 1088.00 | AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug | 8B0001 | Signaling Intermediate Ab | 重组抗体 | 2828.00 | AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug | 8A0022 | Phospho-Specific Ab | 受体 | 2828.00 | AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug | 16440 | Fluorescent Anti-IgGs | 荧光染料 | 1088.00 | AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug | 16518 | Fluorescent Anti-IgGs | 抗小鼠 | 4278.00 | AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug | 16900 | Streptavidin Conjugates | 其他生物染料 | 1378.00 | AAT Bioquest/GCNT3 Antibody/8C14708/50 ug | 8C14708 | Signaling Intermediate Ab | 功能性抗体 | 2828.00 | AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate | 12518 | Reporter Gene Enzymes | 其它检测试剂盒 | 2103.00 | AAT Bioquest/ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 &mi | 1100 | mFluor™ Dyes and Kits | 标记试剂盒 | 2103.00 | AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests | 10058 | Diagnostic Molecules | 定量试剂盒 | 2828.00 | AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each | 38101 | cAMP GPCR Assays | 常规细胞株 | 0.00 |