Description:BPI,Human,ELISAkitTheantimicrobialproteinBPI(BactericidalPermeABIlityIncreasingprotein)isa55kDaproteinfoundintheprimary(azurophilic)granulesofhumanneutrophilsandhasalsobeendetectedonsurfaceofneutrophils,smallintestinalandoralepithelialcells.BPIisabactericidalcompoundthatispresentinpolymorphonuclearcells(PMN)andinlowerlevelsinthespecificgranulesofeosinophils.BPIpossesseshighaffinitytowardthelipidAregionoflipopolysaccharides(LPS)thatcomprisetheouterleafletofthegram-negativebacterialoutermembrane.BindingofBPItothelipidAmoietyofLPSexertsmultipleanti-infectiveactivitiesagainstgram-negativebacteria:1)cytotoxicityviasequentialdamagetobacterialouterandinnerlipidmembranes,2)neutralizationofgram-negativebacterialLPS,3)opsonizationofbacteriatoenhancephagocytosisbyneutrophils.AirwayepithelialcellsconstitutivelyexpresstheBPIgeneandproducetheBPIproteinand,therefore,BPImaybeacriticaldeterminantinthedevelopmentofLPS-triggeredairwaydisease.InflammationinducedbyLPSpossIBLycontributestothedevelopmentofrapidairflowdecline,aseriousandoftenfatalcomplicationofhematopoieticcelltransplantation.FurThermore,a21kDabioactiverecombinantfragmentofBPI,rBPI21,wasshowntoconferasurvivaladvantageagainstinvasivepneumococcaldiseasebybindingtothegram-positivebacterialpathogen,pneumolysin.InplasmaofhealthyindividualsBPIispresentatlevelsofSpecifications:CatalognumberHK314-02ProducttypeAssaysQuantity2x96det.Standardrange100-25,000pg/mlDetectionlevel250pg/mlWorkingvolume100µl/wellSpeciesHumanApplicationThehumanBPIELISAkitistobeusedfortheinvitroquantitativedeterminationofhumanBPIincellculturemedium,plasma,woundfluidandbronchoalveolarlavagefluid.DiseaseInfectiousdiseasesPrincipleThehumanBPIELISAisaready-to-usesolid-phaseenzyme-linkedimmunosorbentassaybasedonthesandwichprinciplewithaworkingtimeof4½hours.Theefficientformatof2plateswithtwelvedisposable8-wellstripsallowsfreechoiceofbatchsizefortheassay.Samplesandstandardsarecapturedbyasolidboundspecificantibody.BiotinylatedtracerantibodywillbindtocapturedhumanBPI.Streptavidin-peroxidaseconjugatewillbindtothebiotinylatedtracerantibody.Streptavidin-peroxidaseconjugatewillreactwiththesubstrate,tetramethylbenzidine(TMB).Theenzymereactionisstoppedbytheadditionofoxalicacid.Theabsorbanceat450nmismeasuredwithaspectrophotometer.Astandardcurveisobtainedbyplottingtheabsorbance(linear)versusthecorrespondingconcentrationsofthehumanBPIstandards(log).ThehumanBPIconcentrationofsamples,whicharerunconcurrentlywiththestandards,canbedeterminedfromthestandardcurve.StorageandstabilityProductshouldbestoredat4°C.Underrecommendedstorageconditions,productisstableforoneyear.PrecautionsForresearchuseonly.Notforuseinoronhumansoranimalsorfordiagnostics.Itistheresponsibilityoftheusertocomplywithalllocal/stateandFederalrulesintheuseofthisproduct.HycultBiotechisnotresponsibleforanypatentinfringementsthatmightresultwiththeuseorderivationofthisproduct.References1.Espinoza,Jetal;Antimicrobialpeptidesinamnioticfluid:defensins,calprotectinandbacterial/permeability-increasingproteininpatientswithmicrobialinvasionoftheamnioticcavity,intra-amnioticinflammation,pretermlaborandprematureruptureofmembranes.JMaternFetalNeonatalMed2003,13:22.Maris,Netal;Antiinflammatoryeffectsofsalmeterolafterinhalationoflipopolysaccharidebyhealthyvolunteers.AmJRespirCritCareMed2005,172:8783.Gubern,Cetal;Naturalantibioticsandinsulinsensitivity,theroleofbactericidal/permeability-increasingprotein.Diabetes2006,55:2164.Manco,Metal;EffectofmassiveweightlossoninflammatoryADIpocytokinesandtheinnateimmunesysteminmorbidlyobesewomen.JClinEndocrinolMetab2007,92:4835.Strunk,Tetal;Reducedlevelsofantimicrobialproteinsandpeptidesinhumancordbloodplasma.ArchDisChildFetalNeonatalEd2009,94:F230