Application of Intracellular Alkaline Phosphatase Activity Measurement in Detection of Neutrophil Adherence In Vitro英文文献资料.docVIP

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Application of Intracellular Alkaline Phosphatase Activity Measurement in Detection of Neutrophil Adherence In Vitro英文文献资料.doc

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Application of Intracellular Alkaline Phosphatase Activity Measurement in Detection of Neutrophil Adherence In Vitro英文文献资料

HindawiPublishingCorporation MediatorsofIn?ammation Volume2006,ArticleID19307,Pages1–8 DOI10.1155/MI/2006/19307 ResearchCommunication ApplicationofIntracellularAlkalinePhosphataseActivity MeasurementinDetectionofNeutrophilAdherenceInVitro KatarzynaBednarska,MagdalenaKlink,andZo?aSulowska DepartmentofExperimentalImmunology,CentreforMedicalBiology,PolishAcademyofSciences,Lodowa106,Lodz93-232,Poland Received28April2006;Accepted13July2006 Wehaveproposedtheuseofthe?uorimetricmethodwith4-methylumbelliferylphosphate(4-MUP)speci?csubstrateforthe alkalinephosphatasedeterminationintheneutrophiladhesionassay.Weprovideevidencethattheendogenousneutrophilalkaline 4 6 phosphatase(NAP)activityevaluationisreliabletoquantifyneutrophiladhesionatawiderangeofcellnumbers(10 –10 ).The resultsobtainedby?uorimetricNAPactivitytestcorrelatetotheresultsofadherenceevaluatedusingtheMTTreductionassay. The?uorimetricNAPactivitytestmaybeappliedforrestingaswellasactivatedneutrophilswithouttheriskoftheactivators interferencesintothetest.Thealkalinephosphatasesurveywiththeuseof4-MUPsubstrateisrecommendedhereinasasensitive, repeatable,simple,andreliablemethodoftheneutrophiladherencedeterminationinvitro. Copyright?2006KatarzynaBednarskaetal.ThisisanopenaccessarticledistributedundertheCreativeCommonsAttribution License,whichpermitsunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalworkisproperly cited. INTRODUCTION some?uorescentdyeslike?uoresceinorcalceinderivatives requireprelabelingofthecellswhichmayresultincytotoxic e?ects[11]. Polymorphonuclear neutrophils (PMNs) are implicated in anin?ammatoryresponseofthehost.Theadhesionofneu- trophilstothevesselwallandextracellularmatrixproteins isrequiredforneutrophiltransendothelialmigrationtothe sitesofin?ammation[1]. Theapplicationof?uorogenicsubstratesfordetectionof intracellularphosphatasesactivityhasbeenproposedasthe method that helps to overcome the above mentioned lim- itations [12]. Phosphatasesarewidely distributed e