Description:

REAADSAnti-Cardiolipin(aCL)IgG/IgM is an ELISAtestkitforthedeterminationof IgG/IgManti-cardiolipin antibodiesinhumanserumorplasma inindividualswithsystemiclupus erythematosus(SLE)andlupus-likedisorders(anti-phospholipidsyndrome).

KitComposition:

Reagents,StorageandStABIlity

• Stabilizedbeefheartcardiolipin(diphosphatidylglycerol)coatedmicrowells:96(12stripsof8breakawaywells),withframe.
• SampleDiluent:1bottle(60ml)(greensolution),containsbovinecalfserum. Containssodiumazide.
• aCLIgGCalibratorSera(1-high,2-moderate,3-low)(human): 3vials(0.250ml)seeviallabelforantibodyconcentrationinGPLunits.Calibrator2shouldbeusedwhenperformingsinglepointcalibration. Containssodiumazide.
• aCLIgMCalibratorSera(1-high,2-moderate,3-low)(human): 3vials(0.250ml)seeviallabelforantibodyconcentrationinMPLunits.Calibrator2shouldbeusedwhenperformingsinglepointcalibration. Containssodiumazide.
• aCLIgGPositiveControlSerum(human): 1vial(0.250ml)seeviallabelforexpectedGPLrange. Containssodiumazide.
• aCLIgMPositiveControlSerum(human): 1vial(0.250ml)seeviallabelforexpectedMPLrange. Containssodiumazide.
• aCLNormalControlSerum(human): 1vial(0.250ml)seeviallabelforexpectedGPLandMPLranges. Containssodiumazide.
• anti-humanIgG(goat)HRP-ConjugatedAntibodySolution:1bottle(15ml)(bluesolution).
• anti-humanIgM(goat)HRP-ConjugatedAntibodySolution:1bottle(15ml)(redsolution).
• OneComponentSubstrate(TMBandH2O2): 1bottle(15ml)readytouse.
• StoppingSolution: 1bottle(15ml)(0.36Nsulfuricacid).
• WashConcentrate:2bottles(30ml) (33XPBS).

Storeat2–8°C.DoNotFreeze.

MeasurementPrinciple:

ThetestisperformedasanindirectELISA.Dilutedserumsamples,calibratorsera,andcontrolsareincubatedincardiolipincoatedmicrowells,allowingaCLantibodiespresentinthesamplestoreactwiththeimmobilizedantigen.Aftertheremovalofunboundserumproteinsbywashing,antibodiesspecificforhumanIgGorIgMlabeledwithhorserADIshperoxidase(HRP)areaddedformingcomplexeswiththecardiolipinboundantibodies.Twoenzymeconjugatedantibodysolutionsareprovided,onespecificforhumanIgGantibodiesandonespecificforhumanIgMantibodies.TheconcentrationofIgGaCLantibodiesandIgMaCLantibodiesmustbedeterminedseparately.Followinganotherwashingstep,theboundenzyme-antibodyconjugateisassayedbytheadditionoftetramethylbenzidine(TMB)andhydrogenperoxide(H2O2)asthechromogenicsubstrate.ColordevelopsinthewellsatanintensityproportionaltotheserumconcentrationofaCLantibodies.ResultsareobtainedbyreadingtheO.D.(opticaldensityorabsorbance)ofeachwellwithaspectrophotometer.Calibratorseraareprovided,withtheIgGandIgMaCLconcentrationsexpressedinGPLorMPLunits,respectively.TheseunitsaretraceabletotheinternationallyrecognizedpreparationsofthePhospholipidStandardizationLaboratory,UniversityofLouisville.Theuserhastheoptionofrunningeitherasinglepointcalibratororafour-pointcalibrationcurve.Forsinglepointcalibration,dividingtheconcentrationvalueofthecalibratorserabytheO.D.valueofthecalibratorprovidesaconversionfactor(oneforIgGandoneforIgMaCL).TheO.D.valuesofalltheothersamplesaremultipliedbytheconversionfactorstoobtainIgGandIgMaCLantibodyconcentrationsinstandardunits.OneGPLunitisequivalentto1µg/mLofanaffinitypurifiedstandardIgGsample,andoneMPLunitisequivalentto1µg/mLofanaffinitypurifiedstandardIgMsample.Formultipointcalibration,performalinearregressionanalysiswithcalibratorvaluesagainstcalibratorO.D.s.Controlandpatientresultsaredeterminedfromthecalibrationcurve.

KitSizes:

CATALOG#	TYPE	WELLS	FORMAT
K023-001	IgG/IgM	96	rapidformat(15-15-10)
K023-002*	IgG/IgM	288	rapidformat(15-15-10)
K026-001	IgA	96	rapidformat(15-15-10)
K026-006*	IgA	288	rapidformat(15-15-10)
K11139*	IgG	96	extendedincubations(30-30-30)
K12747*	IgG	288	extendedincubations(30-30-30)
K11140*	IgM	96	extendedincubations(30-30-30)
K12748*	IgM	288	extendedincubations(30-30-30)
K11141*	IgA	96	extendedincubations(30-30-30)
K12770*	IgA	288	extendedincubations(30-30-30)

*Thisproductmayrequireaspecialorder.Pleaseinquireatinfo@Diapharma.comor1-800-526-5224.

Background:

Anti-cardiolipin(aCL)antibodiesareassociatedwiththepresenceofbothvenousandarterialthrombosis,thrombocytopenia,andrecurrentfetalloss.Theseautoantibodiesarefrequentlyfoundinpatientswithsystemiclupuserythematosus(SLE)andotherautoimmunediseases,aswellasinsomeindividualswithnoapparentpreviousunderlyingdisease.

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