Description:
RoxFIX-A isachromogenicassaykitforquantitativedeterminationofhumanFactorIXa(FIXa)contaminationinhumanFactorIX(FIX)concentrates.
ThekitmethodcomprisesactivationofhumanFXbysampleFIXainthepresenceofFVIII,thrombin,calciumionsandphospholipids.GeneratedFXaisdirectlyproportionaltotheFIXaactivityinthesampleandismeasuredthroughhydrolysisofachromogenicFXasubstrate.
- ChromogenickitbaseduponFXageneration.
- Quantificationlimit=0.02mIU/mlFIXa.
- Detectsapreactivationlevelintheorderof0.002%.
- Reconstitutedreagentsarestablefor72hat2-8°C.
KitComposition:
- Reagent1(2vials)
Reagent1containslyophilizedFactorVIIIandhumanFX.Reconstitutewith2.8mlwater.Allowtostandfor5minat20-25°Cwithintermittentgentlemixingforcompletereconstitution. - Reagent2(2vials)
Reagent2containslyophilizedthrombin,calciumchlorideandphospholipids.Reconstitutewith4.0mlwater.Allowtostandfor5minat20-25°Cwithintermittentgentlemixingforcompletereconstitution. - FXaSubstrate,5ml(1vial)
LiquidsolutionofchromogenicFXasubstrate(Z-D-Arg-Gly-Arg-pNA),2.5mmol/l,containingathrombininhibitor.Readytouse. - FIXaDiluentBuffer,StockSolution,20ml(1vial)
Liquidstocksolutionofdiluentbuffer.Beforeuse,dilute1+9withwatertoobtaina0.05mol/lTris-HClbufferworkingsolution,pH7.5with1%bovineserumalbumin.
Note:AllreconstitutionsanddilutionsshouldbemadewithwaterofaqualityofatleastNCCLSTypeIIwater.
MeasurementPrinciple:
FIXaactivityinFIXconcentratesisdeterminedinachromogenicmethod,inwhichhumanFXisactivatedbycontaminatinghumanFIXaintheFIXconcentrateinthepresenceofFVIII,thrombin,calciumionsandphospholipid.TheamountofgeneratedhumanFXaisdeterminedfromthehydrolysisofachromogenicFXasubstrate.ThesampleFIXaactivityisdeterminedbythesloperatiomodelinwhichthepotencyofthesampleiscalculatedvs.aFIXastandardwithpotencyexpressedinInternationalUnits(IU).
