Description:

Technozym®Glu-PlasminogenisahighlysensitivecompletesandwichELISAfortheselectivedeterminationofGlu-Plasminogeninhumanplasma.TheGlu-PlasminogenELISAkitisbasedontwomonoclonalantibodies;acatchingantibodyandadetectingantibodyspecificfortheGlu-formofPlasminogen. TheGlu-plasminogenELISAisusefulforthedeterminationofGluplasminogenlevels,particularlyinstudiesregardingthroxmboembolicresearch,lyticresearchorinresearchstudiesofhyperfibrinolysis.

Specificity:

ThemonoclonalantibodiesemployedinthistestsystemrecognizeonlyuncleavedGlu-plasminogen.Thus,theresultsarenotaffectedbythepresenceofplasmin-alpha-2-antiplasmincomplexesorplasmin-modifiedLys-plasminogen.TheassaymeasuresGlu-plasminogeninarangefrom0.06-0.5micrograms/milliliter.Normalplasmalevelsare60-250micrograms/milliliter.Theinter-andintra-assayvariationsarelessthan10%and5%,respectively.

KitComposition:

KitComposition

1. Plate+PlateCover
   12x8wellmicrotitrestripsprecoatedwitha monoclonalanti-Plasminogenantibodyandblocked with1%bovineserumalbumin(BSA),lyophilized.
   (TC-CodeGX)
2. Standard
   1xlyophilizedNormalPlasma
   (TC-CodeBJ)
3. POX-Antibody
   1xconjugatedpolyclonalantiPlasminogenantibodies(concentrated).
   (TC-CodeBG)
4. IncubationBuffer
   (PBS;pH7,3);containsstABIlizerprotein;0.05%proclin;and bluedye,1bottle,90ml,readyforuse
   (TC-CodeNB)
5. Substrate–(greencap)
   1x12mlTMB(Tetramethylbenzidine)insubstratebuffercontaining H₂O₂.Readytouse.
   (TC-CodeKN)
6. StopSolution–(redcap)
   1x12ml0.45mol/lSulphuricAcid
   (TC-CodeKK)
7. WashingBufferConcentrate-(1+11,5)
   (PBS;pH7,3)containingdetergent;0,01%merthiolat,1bottle,80ml
   (TC-CodeNA)

Kitstorage:Storeallcomponentsat2-8°C.

MaterialsRequired(butnotsupplied)

1. MicroPipettesandamultichannelmicropipette;pipettetips.
2. Glassorplastictesttubesfordilutingthestandard+samples.
3. Laboratorybottlesorbeakersandgraduatedcylindersfordilutingwash andincubationbuffer.
4. Distilledordeionizedwater.
5. Absorbentpapertowels.
6. Microtitreplatewasher(alternatively,washingcanbeperformed manuallyusingamultichannelpipetteorrepeatingsyringe).
7. Amicrotitreplatereaderequippedwitha450nmfilterand,ifpossIBLe,a 620nmreferencefilter.
8. A37°Cincubator.
9. Graphpaper.

MeasurementPrinciple:

TheTCGlu-plasminogenELISAtestisasolidphaseenzyme immunoassay.

Background:

Plasminogenistheinactiveprecursorofplasmin,thecentralenzyme responsibleforfibrinolysis.Plasminogen,whichissynthesizedbytheliver, isa92,000daltonsingle-chainglycoprotein.Itcirculatesinplasmaata concentrationofapproximately200µgpermlwithahalflifeof2.2days. Throughtheactionofcertainproteasessuchastissue-typeplasminogen activatororurokinase,asinglepeptidebondintheplasminogenmolecule iscleavedtoyieldthetwochainmolecule,plasmin.Plasmin’smajor functionisphysiologicclotdissolutionthroughthedegradationoftheinsoluble polymerfibrinintosolublefragments.Thus,theconcentrationof plasminogenisoneofthenumerousfactorscriticallyinfluencingtherateof fibrinolysisinvivo.

Plasminogenlevelsareoftendepressedinbothacuteandchronichepatic disease.Thismaybedueeithertodecreasedsynthesisorincreased consumptionduringdisseminatedintravascularcoagulation.Inprimary biliarycirrhosisorearlycommonbileductobstructionplasminogenvalues arenormalorevenelevated.Abnormallylowplasminogenvalueshave beenfoundinpatientswithgeneralizedhyperfibrinolysisandneonateswith Lipströmsyndrome.