Description:
Technozym® Fibronectinisa highlysensitivecompletesandwichELISAkitfortheselectivedeterminationofintactfibronectininhumanplasma.TheFibronectinELISAkitisbasedontwomonoclonalantibodies.Onemonoclonalantibodyisimmobilizedtomicrotitreplatesforuseasacatchingantibody,thesecondperoxidased-labeledmonoclonalantibodyrecognizesexclusivelyuncleavedfibronectin.
TheTechnozym®Fibronectinisusedtomeasurehumanplasmafibronectinlevels,particularlyinresearchsamplesfromseveretrauma,shock,sepsis,hepaticdiseaseorclottingdisorders.
Specificity:
Theperoxidaseconjugatedanti-fibronectinmonoclonalantibodyusedinthekitrecognizesonlyintactfibronectin,i.e.splitproductsofthemoleculearenotdetected.Innormalplasma,approximatelytwo-thirdsofthetotalquantityofcirculatingfibronectinisinintactform.Inpatientswithseveresepticemiaordisseminatedintrovascularcoagulation(DIC)fibronectinsplitproductsaregeneratedbytheactionofleucocyteelastaseorplasminontheintactmoleculeleADIngtoarapiddecreaseinthelevelsoffunctionalityintactfibronectin.Thenormalplasmaconcentrationoftotalfibronectinis330±80μg/ml,andnativeuncleavedfibronectinis70–148 μg/ml.
KitComposition:
- Plate+PlateCover
12x8wellmicrotitrestripsprecoatedwithamonoclonalanti-FNcoatingantibodyinbicarbonatebuffer,0.01%ThimerosalandblocedwithStABIlcoat
(TC-CodeGW) - Standard
1xlyophilizednormalpooledhumanplasma.
(TC-CodeAJ) - POX-Antibody
1xconjugatedmonoclonalantiFNantibody(concentrated).
(TC-CodeAG) - DilutionBuffer–(whitecacp)
3x20m.2.5xconcentrated(PBS,1%BSA)
(TC-CodeDS) - Substrate–(greencap)
1x12mlTMB(Tetramethylbenzidine)insubstratebuffercontainingH2O2.ReadyUse.
(TC-CodeKN) - StopSolution–(redcap)
1x15ml0.5mol/lsulphuricacid
(TC-CodeKK) - WashBuffer–(bluecap)
1x20ml.12.5xconcentrated(PBS0.5%,Tween20)
(TC-CodeBE)
MeasurementPrinciple:
TheTechnozym® Fibronectintestisasolidphaseenzymeimmunoassay.
Background:
Plasmafibronectinisahighmolecularweightglycoproteincomposedoftwonearlyidenticalpolypeptidechains(each220kD).Itissynthesizedandsecretedbytheliverandcirculatesataconcentrationofapproximately330μg/mlplasma.
Fibronectinbelongstothegroupof“cellattachmentproteins”.Itsdimericstructureallowsittofunctionasamolecularglue,holdingvariousmoleculestogetherthroughitsbindingdomains.Principalbindingdomainsexistforfibrin,heparin,Staphyloccusaureus,collagenandthecellsurface.Fibronectinhasbeenshowtoplayaroleinfibrincoltformation,plateletfunction,fibrinolysis,chemotaxis,phagocytosisandopsinization.
Loweredplasmafibronectinlevelsareassociatedwithsepticemia,traumaandarealsoobservedduringthepost-operativeperiod.
Hepaticdisordersarealsooftenassociatedwithreducedfibronectinlevels.FibronectinbindstofibrinthroughfactorXIIIandlevelsmaybereducedbyactivationoftheclottingsystem.
Fibronectinisan“acutephaseprotein”anditslevelmaybeelevatedduringtheacutephaseandcomplicationsduringpregnancy.Certainmalignanciesareassociatedwithhighfibronectinlevelsandthisproteinmayplayaroleincellmetastasis.
