Description:

Technozym®t-PAAntigenEDTAELISAisa highlysensitivecompletesandwichELISAkitforthequantitativedeterminationofTissue PlasminogenActivator(tPA)inhumanplasma.TheassayisadoublesandwichELISAemployingtwomonoclonalantibodies.ThesystemdetectsbothcomplexedanduncomplexedtPA.Detectionlimitis1ng/mL.Acidifiedcitratedplasma,citrated,CTADorEDTAplasmascanbeused.TheassayisstandardizedwiththeInternationalstandard98/714.Assaytimeis2.5hours.Kitcontentsincludestandards,highandlowcontrols,coatedplate,peroxidase-labeledantibody,buffersandreadytouseTMBsubstrate.ThesamplebuffercontainsEDTAforincreasedrecoveryoft-PAininnormalsamples.

KitComposition:

1. ELISAteststrips(12)with8wellseach,coatedantit-PAmonoclonalantibody;thedryingagentissuppliedinanaluminiumbag.
2. Washingbufferconcentrate(PBS;pH7.3);containingdetergent;0.01%merthiolate;1bottle,80ml.
3. Incubationbuffer(PBS;pH7.3);containsstABIliserprotein;0.05%proclin;andbluedye,1bottle,90ml,readyforuse.
4. Sampledilutionbuffer(PBS,EDTA);containsstabiliserprotein;1bottle,20mL
5. Calibrators(Standards)numbered;lyophilised;1bottleeach.Concentrationsarelot-dependent;consultlabelonthevial.
6. Controlplasmas“lowlevel”and“highlevel”forcheckingpurposeslyophilised;1bottleeach.Concentrationsarelot-dependent;consultthelabelonthevial.
7. ConjugatemonoclonalAnti-t-PAPOX;dyedblue;1bottle,0.3ml.
8. ChromogenTMB(tetramethylbenzidine);1bottle,12ml;readyforuse.
9. Stoppingsolutionsulphuricacid0.45mol/l;1bottle12ml;readyforuse.
10. Adhesivefilm:forELISAteststrips(2).

MATERIALREQUIRED(butnotsuppliedwiththekit)

1. Distilledwater
2. Testtubesfordilutingstandardandsamples
3. Measuringcylinder(100mLand1000ml)
4. PrecisionPipettes(10,100and1000μl)
5. Variablepipette(1000μl)
6. Multichanneland/ordispensingpipettes(100and200μl)
7. ELISAwasherormultichannelpipette
8. ELISAreaderwith450nmfilter,witha620nmreferencefilterifavailable.
9. Incubator(37°C)

Background:

REFERENCERANGE

Normalrangefort-PAantigenisbetween2-8ng/mlItisrecommendedthatindividuallaboratoriesestablishtheirownranges.Forplasmaobtainedaftervenousocclusionvaluesgreaterthan+10ng/ml,shouldbeobtained.
Afailureoft-PAlevelstoincreaseuponvenousocclusionisindicativeofanimpairedfibrinolyticcapacityandmayindicatethrombotictendencies.

STANDARDIZATION

ThecalibrationmaterialusedistheWHOinternationalstandardfortissueplasminogenactivator(t-PA).