Application:
StripWesternBlotforforre-probingandformassspectrometry

Features:
-Stripblotin5~15minutes
-Incubationatroomtemperature
-Nostenchsmellingwithoutadditionof2-mercaptoethanoloritsanalogs
-Aproprietyformulationwithaneco-friendlyandnon-hazardousorganicsolvent(withlightodor)includedtocompletelydisassociatetheboundantibodies

ApplicationI.StripWesternBlotforRe-probing
AComparisonofBlotFresh™StrippingReagentwithPierce"sRestore™BlotStrippingBuffer

-BlotStrippedandRe-probedwiththeSameAntibody*:

TotalAkt(catalog#A0501-1)withdifferenttreatmentsinNIH3T3cells
*Develop2minuteswithECLPlus™andexpose20secondsonKodaknegatives

-BlotFresh™StrippingReagentisMoreEffectiveinBreakingDowntheAntigen-AntibodyComplexthanPierce"sRestore™StrippingBuffer:

BlotFresh™StrippingReagent*Pierce"sRestore™StrippingBuffer*
     Original          Exposex1"    Exposex5"         Original        Exposex1"    Exposex5"

*Theblotswereinitiallyprobedwithβ-actinantibodywithexposurefor10seconds.ThentheblotswerestrippedwithBlotFresh™StrippingReagentfor5minutesandwithPierce"sRestore™StrippingBufferfor15minutesrespectively.Afterwashes,thestrippingeffectwastestedbydevelopmentofblotswithECLPlus™for2minutesfollowedbyexposuretoKodakBioMaxfilmsfor1and5minutesrespectively

-BlotFresh™StrippingReagentStripsaBlotfollowedbyRe-probingwithaDifferentAntibody:


A.BlotwasprobedwithantibodyagainsttotalAkt(catalog#A0501-1)*.
B.BlotwasstrippedwithBlotFresh™StrippingReagentandre-probewithantibodyagainstphospho-Akt(catalog#A0503-1)*.
*Lane1isserumstarvedovernightwhilelane2~5weretreatedwithPDGFindifferentconcentrationsinNIH3T3cells.Theblotsweredeveloped2minuteswithECLPlus™reagentandexpose10secondsonKodaknegatives(ECLPlus™isatrademarkofGEHealthcare-Amersham)

ApplicationII.StripWesternBlotforMassSpectrometry
ProceduresforStrippingWesternBlotforMassSpectrometry:
-Stripblotfor15minutesatRTtoremoveprimaryandsecondaryantibodies
-Locatetheproteinbycomparingthestrippedblotandexposedfilmandcutthebandfromblot
-Digestthecutbandfollowedbymassspectrometry(seethefollowingpicture)

DataSheet

Testimonials:
...hadreceivedsampleofBlotFresh™WesternBlotStrippingReagent.Fantasticproduct!!Haveusedtostripsameblot2x,andresultingthreerADIographsareindistinguishablefromprimaryblotswiththesecondtwoantibodies.I"llbeorderingsometoday...
------byDr.CatherineBair,NCI,NIH

Icannotlivewithoutit!Itisjustamazing.IusedtousePierce’sandIoftenneedtore-runWesternduetodirtyblotafterstrip.SinceIstartedusingthisproduct,mylifehasbeenchanged…ItmakesmyresearcheasierandIdonotneedtore-rungelanymore…Andthestrippingprocessisjust2minutes.Itisreallyamazingproduct!
------byDr.JenniferLin,theGeorgeWashingtonUniversity

SomeProductCitations:
1.MahmoudM.El-Mas,MingFan,andABDelA.Abdel-Rahman.Endotoxemia-MediatedInductionofCardiacInducIBLeNitric-OxideSynthaseExpressionAccountsfortheHypotensiveEffectofEthanolinFemaleRats.J.Pharmacol.Exp.Ther.2008;324:368-375
2.GrzegorzWoszczek,Li-YuanChen,SahrudayaNagineni,SaraAlsAATy,AnyaHarry,CaroleaLogun,RafalPawliczak,andJamesH.Shelhamer.IFN-gammaInducesCysteinylLeukotrieneReceptor2ExpressionandEnhancestheResponsivenessofHumanEndothelialCellstoCysteinylLeukotrienes.TheJournalofImmunology,2007;178:5262-5270
3.DavidABaker,AricMadayag,LarsVKristianse,JamesHMeador-Woodruf,VahramHaroutuniaandIlangovanRaju.ContributionofCystine–GlutamateAntiporterstothePsychotomimeticEffectsofPhencyclidine.Neuropsychopharmacology,2007;Aug.29onlinepublication
4.AshleyB.WilliamsandPatriciaL.Foster.TheEscherichiacoliHistone-likeProteinHUHasaRoleinStationaryPhaseAdaptiveMutation.Genetics.2007;177:723–735