Description:
REAADSAnti-Cardiolipin(aCL)IgAis an ELISAforthedeterminationofIgAanti-cardiolipin antibodiesinhumanserumorplasma inindividualswithsystemiclupus erythematosus(SLE)andlupus-likedisorders(anti-phospholipidsyndrome).
Advantages:
- SpecificdeterminationofIgAaCLantibodies
- ConvenientELISAprocedure
- Objective,accurateandreproducIBLe
- Reagentcompletekit
- Shortincubationsatroomtemperature
KitComposition:
Reagents,StorageandStABIlity
- Stabilizedbeefheartcardiolipin(diphosphatidylglycerol)coatedmicrowells:96(12stripsof8breakawaywells), withframe.
- SampleDiluent: 1bottle, 60ml(greensolution),containsbovinecalfserumand sodiumazide.
- aCLIgACalibratorSerum: 3vials, 0.250ml(1-high,2-moderate,3-low)(human),seeviallabelforantibody concentrationinAPLunits.Calibrator2shouldbeusedwhenperformingsinglepointcalibration.Containssodiumazide.
- aCLIgAPositiveControlSerum: 1vial,0.250ml(human),seeviallabelforexpectedAPLrange.Containssodiumazide.
- aCLNormalControlSera: 1vial, 0.250ml,(human),seeviallabelforexpectedAPLrange.Containssodiumazide.
- anti-humanIgA(goat)HRP-conjugatedantibodysolution:1bottle,15ml(orangesolution).
- OneComponentSubstrate: 1bottle, 15ml(TMBandH2O2),readytouse.
- StoppingSolution:1bottle, 15ml(0.36Nsulfuricacid).
- WashConcentrate2bottles, 30ml (33XPBS).
Storeat2–8°C.DoNotFreeze.
PrincipleandProcedure:
Principle
ThetestisperformedasanindirectELISA.Dilutedserumsamples,calibratorsera,andcontrolsareincubatedin cardiolipincoatedmicrowells,allowingaCLantibodiespresentinthesamplestoreactwiththeimmobilizedantigen. Aftertheremovalofunboundserumproteinsbywashing,antibodiesspecificforhumanIgAlabeledwith horserADIshperoxidase(HRP)areaddedformingcomplexeswiththecardiolipinboundantibodies.Following anotherwashingstep,theboundenzyme-antibodyconjugateisassayedbytheadditionoftetramethylbenzidine (TMB)andhydrogenperoxide(H2O2)asthechromogenicsubstrate.Colordevelopsinthewellsatanintensity proportionaltotheserumconcentrationofIgAaCLantibodies. ResultsareobtainedbyreadingtheO.D.(opticaldensityorabsorbance)ofeachwellwithaspectrophotometer. Calibratorseraareprovided,withtheIgAaCLconcentrationexpressedinAPLunits.Theuserhastheoptionof runningeitherasinglepointcalibratororafour-pointcalibrationcurve.Forsinglepointcalibration,dividingthe concentrationvalueofthecalibratorbytheO.D.valueofthecalibratorprovidesaconversionfactor.TheO.D. valuesofthecontrolsandpatientsamplesaremultipliedbytheconversionfactortoobtainIgAaCLvalues, expressedinAPLunits.Formultipointcalibration,performalinearregressionanalysiswithcalibratorvalues againstcalibratorO.D.s.Controlandpatientresultsaredeterminedfromthecalibrationcurve.
Procedure
Dilutedpatientserum(preferred)orplasmaisincubatedincoatedmicrowells.Cardiolipinantibodiespresentinthesamplewillbindtothecoatedwells.Afterwashingtoremoveunboundplasmaproteins,enzymeconjugatedanti-humanimmunoglobulinspecificforIgAisadded.Thewellsarewashedagain,andachromogenicsubstrateisadded,resultinginacoloredproduct.Thereactionisstoppedbytheadditionofaweakacidsolution,andthecoloredproductismeasuredinaspectro-photometerat450nm.Testresultsareavailableinlessthanonehour.ValuesforaCLantibodiesarereportedinGPL,MPL,andAPLunits,withassaycutoffsestablishedat23GPL,11MPL,and22APL.
ClinicalPerformance*:
ClinicalSpecificity:Assaycutoffswerechallengedwithahealthyblooddonorpopulation.Usingthestatedcutoffs,theassayswere97%specificforIgG,96%specificforIgM,and95%specificforIgA.
UnselectedSLEpopulationsweretestedtodetermineassaysensitivity;21%ofthesampleswerepositiveforIgG,10%forIgMand26%forIgAantibodieswiththeREAADSaCLassays.TheclinicalsensitivityforthrombosisandthrombocytopeniawasdeterminedbycomparingaCLtestresultsfromtwogroupsofselectedSLEpatients:Group1?withaclinicalhistoryofthrombosisand/orthrombocytopenia;andGroup2?withnohistoryofthrombosisorthrombocytopenia(control).Theresultsareshowninthechartsbelow:
Group1:SLE+Thrombosisand/orthrombocytopenia | IgGaCL | IgMaCL | IgAaCL |
Averagevalue | 30GPL | 9 MPL | 32APL |
%positive | 45% | 25% | 79% |
Group2:SLEControl | IgGaCL | IgMaCL | IgAaCL |
Averagevalue | 8 GPL | 3 MPL | 15 APL |
%positive | 0% | 0% | 0% |
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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2.NaHCO3,Ba(OH)3,H2SO4
3.HCL,NaAlO2,NaHSO4
4.Ca(OH)2,Na2CO3,BaCO3
谢谢了
要原因
请问有什么方法检测细胞内活性氧更好,各位有什么经验,及相关试剂盒推荐一下。
A.H2SO4Na2SO4AgNO3BaCl2.
B.NaOHNa2CO3NaHSO4MgCl2
C.CaCl2NaNO3MgSO4BaCl2
D.HNO3KOHKClK2CO3
请问下有无同学需要H37RA的?我是做EAE模型的,上个月购买了BDDifco公司的H37RA(货号),因为购买的时候只能整盒6支购买,但我们用不了那么多,所以想问问有无同学需要的,100mg/支,800元/支或用等价试剂交换。地址广州。有需要的请私信,谢谢!
(1)优级纯试剂 亦称保证试剂,为一级品,纯度高,杂质极少,主要用于精密分析和科学研究,常以GR表示。
(2)分析纯试剂 亦称分析试剂,为二级品,纯度略低于优级纯,杂质含量略高于优级纯,适用于重要分析和一般性研究工作,常以AR表示。
(3)化学纯试剂 为三级品,纯度较分析纯差,但高于实验试剂,适用于工厂、学校一般性的分析工作,常以CP表示。
(4)实验试剂 为四级品,纯度比化学纯差,但比工业品纯度高,主要用于一般化学实验,不能用于分析工作,常以 LR表示。
以上按试剂纯度的分类法已在我国通用。根据化学工业部颁布的“化学试剂包装及标志”的规定,化学试剂的不同等级分别用各种不同的颜色来标志,见表1。
表1 我国化学试剂的等级及标志
bhclna2so4nano3na2co3
chclnaohna2co3nacl
dba(oh)2nahco3alcl3nahso4