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.
