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商品描述
Description:DxFecalCalprotectinIVDELISAkit
Calprotectin,alsoknownasS100A8/A9heterocomplex,isformedoutofthecalcium-binding,migratingproteinsS100A8andS100A9.Calprotectinisacalcium-andzinc-bindingproteincomplexcomposedof8and14kDsubunitsandcomplexformationiscalciumdependent.Neutrophilsarethemainproducersofcalprotectin,butexpressionhasalsobeenfoundinmonocytes,macrophagesandilealtissueeosinophils.Altogether,S100A8andS100A9proteinscontributetoapproximately40–60%ofthesoluble,cytosoliccontentofgranulocytes.Calprotectinhasantibacterial,antifungal,immunomodulatingandantiproliferativeeffects.Elevatedconcentrationsofcalprotectincanbemeasuredamongothersinplasma,synovialfluid,urineandfeces.Thepresenceofcalprotectininfecesquantitativelyrelatestoneutrophilmigrationtowardsthegastrointestinaltract.CalprotectinisthereforeconsideredareliablebioMarkerforintestinalinflammationthroughoutthegastrointestinaltract.Calprotectininfecesisstableforseveraldaysandevenlongerat4°C.ConsistentlyhigherfecalcalprotectinlevelshavebeenreportedinbothadultandpediatricpatientswithIBDcomparedtoIBSorhealthycontrols.Raisedlevelsoftheproteinindicatepresenceofluminalwhitecells.Suchfindingindicatesthattheintestineisinflamed,althoughthetypeofinflammationisnotdefined.Nowadays,itiscommonlythoughtthatconcentrationsof<50=""μg/g=""represent=""normal,=""50-150=""μg/g=""represent=""a=""weakly=""positive=""test=""and=""above=""these=""values=""indicate=""a=""positive=""test.=""however,=""these=""values=""are=""still=""under=""debate=""and=""might=""be=""laboratory=""dependent.=""in=""most=""cases,=""a=""negative=""calprotectin=""test=""rules=""out=""ibd,=""thereby=""sparing=""most=""people=""with=""ibs=""from=""having=""an=""endoscopic=""procedure.=""the=""elisa=""is=""a=""single=""incubation=""step=""reaction.=""after=""this=""incubation=""the=""wells=""are=""filled=""with=""substrate.=""the=""obtained=""color=""intensity=""is=""proportional=""to=""the=""amount=""of=""calprotectin=""in=""your=""sample.=""the=""amount=""of=""calprotectin=""can=""be=""calculated=""based=""on=""the=""standard=""curve=""of=""calibrated=""native=""calprotectin.=""the=""test=""performance=""is=""assured=""by=""the=""addition=""of=""a=""high=""control=""and=""a=""low=""control.="">
Specifications:
| Catalognumber | DH002 |
|---|---|
| Producttype | Assays |
| Quantity | 1x96det. |
| Standardrange | Measurableconcentrationrangeof16-625μg/g(RangeI;50xsampledilution)or48-1,875μg/g(RangeII;150xsampledilution). |
| Detectionlevel | Theminimumconcentrationwhichcanbemeasuredis16μg/g. |
| Workingvolume | 100µl/well |
| Species | Human |
| Alias | S100A8/S100A9,MRP8/MRP14 |
| Application | TheDxFecalCalprotectinassayisasandwichELISAspecificallydevelopedforquickandreliableinvitroquantificationoffecalcalprotectininstoolsamples.Thisquantificationenablestodiscriminatebetweeninflammatoryboweldisease(IBD)andirritablebowelsyndrome(IBS)andalsoallowsmonitoringofdiseasestateoractivityinIBDpatients.Theresultsofthekitaretobeusedincombinationwithotherlaboratoryandclinicalfindingsforthepurposeofprovidinginformationaboutapathologicalstate.Theanalysisshouldbeperformedbytrainedlaboratoryprofessionals. |
| Disease | Gastroenterology |
| Principle | TheDxFecalCalprotectinELISAisaready-to-usesolid-phaseenzyme-linkedimmunoassaywithaworkingtimeof1hourand30minutes.Theefficientformatof1platewithtwelvedisposable8-wellstripsallowsfreechoiceofbatchsizefortheassay.BiotinylatedcaptureantibodyandHRPconjugateddetectionantibodywillbindtocalprotectinpresentinstandardandsamplesinthemixture.Biotinylatedcaptureantibodywillbindtothestreptavidincoatedplates,incomplexwiththestandardorsamplesandtheHRPconjugateddetectionantibody.HRPconjugateddetectionantibodywillreactwiththesubstrate,tetramethylbenzidine(TMB).Theenzymereactionisstoppedbytheadditionofoxalicacid.Theabsorbanceat450nmismeasuredwithaspectrophotometer.Astandardcurveisobtainedbyplottingtheabsorbance(linear)versusthecorrespondingconcentrationsofthecalprotectinstandards(log).Thecalprotectinconcentrationofsamplesandcontrols,whicharerunconcurrentlywiththestandards,canbedeterminedfromthestandardcurve. |
| Recovery | Recoverywasdeterminedbymixingfecalextractionsampleswithaknowncalprotectinconcentrationindifferentratios.Therecoveryrangedbetween84.9%and116.2%. |
| AutomatedELISAperformance | TheassaycanbeperformedonanautomatedELISAplatformwhichiscapabletoshaketheplateduringincubation.Pleasenotethatshakingthedevicebelowtherecommendedspeedof500rpmwillresultinalowerabsorbance.AssaywastestedonaDynexDS2toevaluatetestperformance.Programforthisplatformisavailableonrequest. |
| PrecisionandReproducibility | Theintra-assayprecisionandreproducibilitywastestedwithextractsoffourfecalsamplesusingtwentyidenticalaliquotsincludingthehighcontrolandlowcontrol.TheCVrangedbetween3.9and5.5%.Todeterminetheinter-assayvariation,fivesamplesweretestedbytwooperatorsbothmanuallyandautomatically(DynexDS2)onthreeseparatedays.TheCVinthissituationranged2.8and9.2% |
| StorageandstABIlity | Productshouldbestoredat4°C.Underrecommendedstorageconditions,productisstableforoneyear. |
| References | 1.Sipponen,Tetal;Fecalcalprotectinindiagnosisandclinicalassessmentofinflammatoryboweldisease.ScandJGastroenterol2015,50:1 2.Henderson,Petal;ThediagnosticaccuracyoffecalcalprotectinduringtheinvestigationofsUSPectedpediatricinflammatoryboweldisease:asystematicreviewandmetaanalysis.AmJGastroenterol2014,109:5 3.Aggio,Retal;Futuremethodsforthediagnosisofinflammatoryboweldisease.DigDis2014,32:4 4.Fengming,Yetal;Biomarkersofinflammatoryboweldisease.DisMarkers2014 5.Stragier,Eetal;TheuseoffecalcalprotectinandlactoferrininpatientswithIBD.Review.ActaGastroenterolBelg2013,76:3 6.Montalto,Metal;Roleoffecalcalprotectiningastrointestinaldisorders.EurRevMedPharmacolSci2013,17:12 |
