商品信息
联系客服

郑重提醒:
无质量问题不接受退换货,下单前请仔细核对信息。
下单后请及时联系客服核对商品价格,订单生效后再付款。
Highlights | FORMULATION:Urokinaseon6%crosslinkedSepharosebeads,providedas50%slurryinpureglycerol. LIGANDDENSITY:0.11mg/mloftheresin. APPLICATIONS:Efficientandconvenientcleavageofplasminogenandotherrecombinantfusionproteinscontainingaurokinase-specificcleavagesite. PROTOCOL:Inordertofindtheoptimumcleavageconditionsforatargetprotein,itisrecommendedtorunpreliminarycleavagereactionsatasmallscale.Successfulcleavagewithurokinaseisdependentuponproperfoldingofthetargetproteinthatenablesaccessoftheurokinaserecognitionsequencebytheenzyme.Onceoptimumcleavageconditionsareobtained,thereactioncanbescaleduptocleavetheentireamountofthetargetprotein.Thetargetproteinshouldbepurifiedtohomogeneityanddialyzedagainst50mMTrisbuffer,0.1MNaCl,pH7.4beforesettingupthecleavagereaction. 1)ResUSPendthebeadsbygentleswirling.Donotvortex. 2)Aliquot250µlofthesuspendedslurryandaddto1mgofthetargetproteininanEppendorftube.Werecommendaproteinvolumeof>400μl/tubetofacilitatepropermixingduringthereaction. 3)Mixgentlybyinvertingthetube(donotvortex)andshakeonarotaryshakerat37°C. 4)Atregulartimeintervals,spindownthetubetoaliquotatestsampleandfreezeitimmediately.Attheendofthereaction,analyzethesamplesbySDS-PAGE. Recoveryofthecleavedtargetprotein: 1)Afterthefusionproteiniscompletelycleaved,spindownthereactionmixturefor2-3minat1,500xg. 2)Removethesupernatantandwashthebeadswith0.2mlof50mMTrisbuffer,0.1MNaCl,pH7.4. 3)Repeatsteps1and2tomaximizetherecoveryofthetargetproteinanditscleavedfragment.Furtherchromatographymaybenecessarytoremovethecleavedfragmentsfromthetargetprotein. |
---|---|
StorageConditions | -20°C |
USAGE | ForResearchUseOnly!NotForUseinHumans. |