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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/ReadiUse™ TCA Deproteinization Sample Preparation Kit/19501/200 Tests
产品编号:19501
市  场 价:¥42060.00
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:$2103.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/ReadiUse™ TCA Deproteinization Sample Preparation Kit/19501/200 Tests
商品介绍
Overview
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Ex/Em(nm)None/None
MWN/A
CAS#N/A
SolventN/A
Storage
CategoryMicroBIOLOGy
FluorescenceImaging
RelatedBiochemicalAssays
Theanalysisofaminoacids,ions,metabolitesandothersmallmoleculesisfrequentlyhinderedbytheinterferenceoflipids,proteinandenzymespresentinbiofluids,cellandtissuelysates.Therefore,deproteinizationisanecessarystepinmanyprocedurespriortoanalysisofbiologicalsamples.AATBioquest"sDeproteinizationSamplePreparationKitoffersarapidmethodforremovingproteinsinbiologicalsamples.Proteinsareprecipitatedbyapplyingtrichloroaceticacid(TCA).Afterremovalofprecipitatedproteins,thepHofthesampleisneutralizedwithneutralizationsolution.TCAbaseddeproteinizationhasbeensuccessfullyandwidelyusedinsamplepreparationpriortoquantitationofsmallmolecules,suchasglycogen,ATP,cAMP,glutathioneandantioxidants.TheTCAmethodprovidedinthiskitcanalsobeusedforremovingproteininlargescale.Biologicalsamplespreparedusingthiskitcanbedirectlyusedinvariousbiochemicalanalysis.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.ProteinPrecipitation:

1.1   Prepareaclearsolutionsamplewithproteinconcentrationlessthan50mg/mLafterhomogenizationandcentrifugation.Keepproteinsamplesonice.

Note:Highproteinconcentrationsamplesmightneedtobediluted.

 

1.2   Add10μLofcoldTCA(ComponentA)into100μLofproteinsampleandvortexbrieflytomixwell.

 

1.3   Keepthesampleonicefor10minutes.Centrifugeat12,000rpmfor5minutes.Transfersupernatant(deproteinizedsample)toanothertube.

Note:ItisrecommendedtoneutralizesamplesinTCA(Step2.1)anduseforrelevantassaysimmediately.However,deproteinizedsamplesinTCAmaybestoredat-70ºCforuptoonemonth.

 

2.TCANeutralization:

2.1   ToneutralizeexcessTCA,add10μLofcoldNeutralizationSolution(ComponentB)intothecollectedsupernatantandvortexbrieflytomixwell.

2.2   Keepthesamplesonicefor5minutes.Samplesarenowdeproteinizedandneutralized.

2.3   (Optional)Readabsorbanceat280nm(OD)tocheckifthesamplesaredeproteinized.

References&Citations
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1.   TuruncE,KaradenizH,ArmaganG,ErdemA,YalcinA.(2013)ElectRochemicaldeterminationofglutathioneinplasmaatcarbonnanotubesbasedscreenprintedelectrodes.CombChemHighThroughputScreen,16,695.

2.   ZhangJ,LuY,ChenD.(2012)[Preparationproceduresofanti-complementarypolysaccharidesfromHouttuyniacordata].ZhongguoZhongYaoZaZhi,37,2071.

3.   XieLY,ZhangY,PengWH,GanBC.(2011)[StudyonmethodsofdeproteinizationfromPhellinusbaumiipolysaccharide].ZhongYaoCai,34,293.

4.   HuangG,ChenY,WangX.(2011)Extractionanddeproteinizationofpumpkinpolysaccharide.IntJFoodSciNutr,62,568.

5.   LiuF,ZhuXH,LiuM,ZhouJ,CheXP,HanRF.(2008)[StudyondeproteinizationanddecolorationinextractionofpolysaccharidesfromRaBDosiarubescens].ZhongYaoCai,31,751.

6.   HaraT,YamakuraF,TakikawaO,HiramatsuR,KawabeT,IsobeK,NagaseF.(2008)Diazotizationofkynureninebyacidifiednitritesecretedfromindoleamine2,3-dioxygenase-expressingmyeloiddendriticcells.JImmunolMethods,332,162.

7.   SamanidouVF,NikolaidouKI,PapadoyannisIN.(2007)DevelopmentandvalidationofanHPLCconfirmatorymethodforthedeterminationofseventetracyclineantibioticsresiduesinmilkaccordingtotheEuropeanUnionDecision2002/657/EC.JSepSci,30,2430.

8.   KataokaM,SetoY.(2003)Discriminativedeterminationofalkylmethylphosphonatesandmethylphosphonateinbloodplasmaandurinebygaschromatography-massspectrometryaftertert.-butyldimethylsilylation.JChromatogrBAnalytTechnolBiomedLifeSci,795,123.

9.   HemmelderMH,deJongPE,deZeeuwD.(1998)Acomparisonofanalyticproceduresformeasurementoffractionaldextranclearances.JLabClinMed,132,390.

10.   KuoTL.(1987)Determinationofparaquatintissueusingion-pairchromatographyinconjunctionwithspectrophotometry.ForensicSciInt,33,177.


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