Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 360/449 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | EnzymeDetection HydrolyticEnzymes |
Related | DiagnosticMolecules BiochemicalAssays |
1.Prepareneuraminidasestandardstocksolution:
Add50µLofddH2OintothevialofNeuraminidaseStandard(ComponentC)tomake2U/mLneuraminidasestandardstocksolution.
Note:Theconcentrationofthisstocksolutionisapproximately2U/mL.TheunusedNeuraminidaseStandardsolutionshouldbedividedintosingleusealiquotsandstoredat-20oC.
2.Prepare200XFluLiteTMBluestocksolution:
Add50µLofddH2OintothevialofFluLiteTMBlue(ComponentA)tomake200Xstocksolution.
Note:TheunusedFluLiteTMBluesolutionshouldbedividedintosingleusealiquotsandstoredat-20oCandkeptfromlight.
3.Prepareneuraminidaseassaymixture:
Add25µLof200XFluLiteTMBluestocksolution(fromStep2)into5mLofAssayBuffer(ComponentB),andmixwell.
4.Prepareserialdilutionsofneuraminidasestandard(0to20mU/mL):
4.1 Add10μLof2U/mLneuraminidasestandardstocksolution(fromStep1)to990µLofassaybuffer(ComponentB)togenerate20mU/mLneuraminidasestandard.
Note:Dilutedneuraminidasestandardsolutionisunstable.Usewithin4hours.
4.2 Take500μLof20mU/mLneuraminidasestandardsolution(fromStep4.1)toperform1:2serialdilutionstoget10,5,2.5,1.25,0.625,0.312and0mU/mLserialdilutionsofneuraminidasestandard.
4.3 Addneuraminidasestandardsandneuraminidase-containingtestsamplesintoasolidblack96-wellmicroplateasshowninTables1and2.
Table1Layoutofneuraminidasestandardsandtestsamplesinasolidblack96-wellmicroplate
BL | BL | TS | TS | …. | …. |
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NA1 | NA1 | …. | …. | …. | …. |
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NA2 | NA2 |
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NA3 | NA3 |
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NA4 | NA4 |
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NA5 | NA5 |
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NA6 | NA6 |
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NA7 | NA7 |
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Note:NA=NAStandards,BL=BlankControl,TS=TestSamples.
Table2Reagentcompositionforeachwell
NAStandard | BlankControl | TestSample |
SerialDilutions*:50μL | AssayBuffer:50μL | 50μL |
*Note:Addtheserialdilutionsofneuraminidasestandardfrom0.312mUto20mUintowellsfromNA1toNA7induplicate.
5.Runneuraminidaseassay:
5.1 Add50μLofneuraminidaseassaymixture(fromStep3)toeachwelloftheneuraminidasestandard,blankcontrol,andtestsamples(seeStep4.3)tomakethetotalneuraminidaseassayvolumeof100µL/well.
Note:Fora384-wellplate,add25μLofsampleand25μLofneuraminidasereactionmixtureintoeachwell.
5.2 Incubatethereactionat37oCorroomtemperaturefor1to2hours,protectedfromlight.
Note:37oCincubationgivesbetterresults.
5.3 MonitorthefluorescenceincreaseatEx/Em=320/460nm(cutoff=420nm)withafluorescencemicroplatereader.
References&Citations | ![]() PrinterFriendlyVersion |
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 |