Formulation	11mMCitrate,50ug/mLCTI(final)
MolecularWeight
Storage	4°C
Purity	N/A
Compound
Assay	N/A
ShelfLife(properlystored)	6months
ChemicalFormula

EffectofCTIontheAPTTandPTofNormalHumanPlasma
ThegraphillustratestheeffectofaddedCTIonthePTandaPTTofnormalhumanplasma.CTIwasaddedtonormalpooledhumanplasmatovariousfinalconcentrations,andthemixtureswereallowedtoincubateforabriefperiodonicepriortoconductingtheassays.ThePTremainsunaffectedwhiletheaPTTdoubleswiththeadditionofapproximately60microgramsofCTIpermlofplasma.

SampleGelInformation:

Gel	Novex4-12%Bis-Tris
Load	CornTrypsinInhibitor,1µgperlane
Buffer	MES
Standard	SeeBluePlus2;Myosin(188kDa),PhosphorylaseB(98kDa),BSA(62kDa),GlutamicDehydrogenase(49kDa),AlcoholDehydrogenase(38kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(17kDa),Lysozyme(14kDa),Aprotinin(6kDa),Insulin,Bchain(3kDa).

Overview:

PurifiedCTI
Corntrypsininhibitor(CTI)isasmallproteinthatislocalizedinthekernelsofmostspeciesofcorn.CTIisnotonlyaninhibitoroftrypsin,butisalsoaspecifichumanfactorXIIainhibitorwhentestedinbloodclottingexperiments(1-6).Theinhibitorformsaone-to-onecomplexwitheithertrypsinorfactorXIIa,andwhenaddedtoplasma,prolongstheactivatedpartialthromboplastintimewithoutaffectingthePTassay(6).ThespecificityforfactorXIIamakestheinhibitorusefulforthesegregationandstudyoftissuefactor(TF)dependentcoagulationreactions(6).

TheuseofCTItostudyTF-dependentreactionshasbeendocumentedinliterature(6,8-10).StudiesindicatethatsuppressionofthecontactpathwayofcoagulationisessentialwhenattemptingtoperformTF-dependentassaysinwholebloodorplasmasamples.TheadditionofCTIatthepointofsamplecollectionpreventsactivationofthecontactpathwayduringsubsequentsampleprocessingsteps,thusreducingin-vitroartifact.ThemostcommonuseofCTIisassociatedwiththrombingenerationassayswhenattemptingtoworkatlowTFconcentrations.HTI’sCTIproductisperformancetestedforthisspecificapplication.

Theproteiniscomprisedof112aminoacidswhichyieldsacalculatedmolecularweightof12,028(7).Thecalculatedmolecularweightandthatwhichhasbeendeterminedbysedimentationequilibriumanalysesofthepurifiedprotein(12,500)areingoodagreement(3).AcomparisonoftheaminoacidsequenceofCTItothatofothertrypsinorserineproteaseinhibitorsrevealslittleornosimilarities(7).

CTIispurifiedusingacombinationofpublishedprocedures(3,4).Tobeginthepurificationprocess,CTIisextractedfromthekernelsoffreshsweetcornintoaphysiologicbuffer.Theextractisthende-fattedusingacetone,andtheproteinisfurtherpurifiedbyemployinggelfiltrationandion-exchangechromatography.ThefinalCTIpreparationappearsasasinglebandbySDS-PAGEanalysesunderbothreducingandnon-reducingconditions.

PreparationsofCTIaretestedfortheABIlitytoprolongtheaPTTassaywithoutaffectingthePTassay.ThespecificactivityofeachlotofCTIisdetermined,andoneunitisdefinedastheamountofCTIrequiredtodoubletheaPTTofnormalhumanplasma.

PurifiedCTIisformulatedin20mMTris,0.15MNaCl,pH7.4andshouldbestoredfrozenat-20°Corcolder.

BloodCollectionTubes
TosimplifytheprocessofcollectingsampleswithaddedCTI,HTIhasdevelopedbloodcollectiontubescontainingCTI.Thesetubes(ourSCAT-27line)simplifytheprocessofconductingTF-dependentstudiesbyallowingyoutodrawblooddirectlyontoananticoagulantcontainingCTI.YoumaychoosetouseourstandardCTI/Citrateformulation(11mMCitrate,50ug/mLCTI)oryoumaycreateyourowncustomformulation.HTI’sbloodcollectiontubesarenotsterileandaremanufacturedandsoldforresearchuseonly.Threestandardsizesareavailablealthoughcustomsizescanbemanufacturedforyou.

SCAT-27-4.5/5-5mLtube

SCAT-27-2.7/5-3mLtube

SCAT-27-1.8/5-2mLtube

 

Properties:

Localization	CornKernals
Modeofaction	formsaone-to-onecomplexwitheithertrypsinorfactorXIIaandinhibitstheirrespectivecatalyticactivity.
Molecularweight	12,500(3)
Extinctioncoefficient	E 1% 1cm,280nm =20.0(determinedbyultracentrifugationstudies)
Structure	singlechainproteincomprisedof112aminoacids(3,7)

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