- TheAdeno-XRapidTiterKitallowsfunctional,hexon-basedadenovirustitration.
- TheAdeno-XqPCRTitrationKitallowsultra-fastadenovirustitrationusingquantitativePCR.
- EachAdeno-XGoStixcassetteisasensitiveinstanttesttodetectadenovirusearlyduringamplificationandtodeterminetheoptimumharvesttime.
- TheAdeno-XRapidTiterKitallowsfunctional,hexon-basedadenovirustitration
- TheAdeno-XqPCRTitrationKitallowsultra-fastadenovirustitrationusingquantitativePCR
- EachAdeno-XGoStixcassetteisasensitiveinstanttesttodetectadenovirusearlyduringamplificationandtodeterminetheoptimumharvesttime
Adeno-XRapidTiterKit
Thiskitisdesignedaroundahexon-specificantibody,whichisusedtolabelinfectedcells.Hexonproteinisencodedbytheadenoviralgenomeandisanessentialcomponentoftheadenoviralcapsidrequiredforadenoviralreplication,butitsexpressiondependsontheE1geneproduct.Thus,E1trans-complementingcelltypessuchasHEK293canbeusedtomeasureinfectiousactivitybecauseonlyinfectedcellswillproducethehexonprotein.Therapidtiterassaytakes1–2hourstosetupandanother3hours(twodayslater)tolabel,stain,andcountinfectedcells.Eachstainedcellcorrespondstoasingleinfectiousunit.
Adeno-XqPCRTitrationKit
TheAdeno-XqPCRTitrationKitprovidesafast,simple,andaccuratemethodfortitratingadenoviralstocksfromallAd5-basedadenoviralvectors,includingrecombinantadenoviruscreatedwithourAdeno-Xexpressionsystems.TheprotocolcombinesqPCRandTBGreenchemistry,allowingyoutodeterminetheviralgenomecopynumberinadenoviralpreparations(i.e.,crudelysatesorpurifiedstocks)fromacalibratedDNAstandardcurve.Thekitdeliversresultsinjust4hours,andbecauseqPCRtitrationissofast,targetcellscanbeinfectedwithaccuratelytitratedvirusonthesamedaythevirusisharvested.
Adeno-XGoStix
Adeno-XGoStixwillrapidly(~10min)tellyouifyouarereadytogowithyouradenovirusprepandisidealfordeterminingthebesttimetoharvestyourvirusfollowingamplification.Simplyapply20µlofculturesupernatanttotheGoStixcassette,chaseitthroughwithfourdropsofbufferandwait2–20minforabandtoappearinthewindowofthecassette.Theamountofadenovirusdetectedinthesupernatantcorrelatesstronglywithadenovirusproductioninsidethecells.Eachtestissensitiveenoughtodetectadenovirusinasmallamountofcrudesupernatantandisalsousedtoensuremaximumyieldsduringtheadenovirusamplificationprocess(weemploythesesametestcassettestodeterminetheoptimumtimetoharvestvirusfrominfectedcells).Just20µlofsupernatantistestedeachdayfollowinginfectionofHEK293cells.Thepresenceofastrongbandinthecassettewindowensuresthathigh-titeradenovirusisharvestedatthepeaktime.