Thisproductisfreezedried.Allwatermoleculeshavebeenremoved.
Everylotistried&testedinarelevantBIOLOGicalassay.
OurBioassay
- Hougaard,C. etal. (2007) BrJ.Pharmacol. 151, 655.
- Chubanov,V. etal. (2012) Br.J.Pharmacol. 166, 1357.
- AlomoneLabsCyPPAenhanceshSK2channelcurrentsintransientlytransfectedHEK293Tcells.Cellswerestimulatedwith100μM CyPPA (#C-110)andcurrentsweremeasuredusingwholecellvoltage-clamp.A.Timecourseat-20mVshowingtheeffectof100μMCyPPAanditsinhibitionwith150nM Apamin(#STA-200).B.Representativecurrentsatcontrolconditions(bottom)andfollowingapplicationof100μMCyPPA(top).Currentswereelicitedbya150msvoltagerampfrom-120to60mVevery10secondsfromaholdingpotentialof-80mV(currentresponsesareshownbetween-120and-10mV).Leaksweresubtractedoffline.
- 1.Kohler,M. etal. (1996) Science 273, 1709.
- 2.Stocker,M. etal. (2000) MolCellNeurosci. 15,476.
- 3.Hougaard,C. etal. (2007) BrJ.Pharmacol. 151, 655.
- 4.Seutin,V. etal. (2007) BrJ.Pharmacol. 151, 568.
- 5.Chubanov,V. etal. (2012) Br.J.Pharmacol. 166, 1357.
Ca2+-activatedpotassiumchannels(KCa)areagroupof6/7-TMionchannelsthatselectivelytransportK+ionsacrossbiologicalmembranes.Theyarebroadlyclassifiedintothreesubtypes:SK,IKandBKchannels(small,intermediateandbigconductance).
Small-conductanceCa2+-activatedK+channels(SKchannels)underliethemediumdurationafterhyperpolarizationthatfollowssingleortrainsofactionpotentialsinmanytypesofneurons.ThisfamilyiscomposedofKCa2.1(SK1),-2.2(SK2),and-2.3(SK3)isoforms,whichareexpresseddifferentiallywithinthecentralnervoussystem(CNS)1,2.
CyPPAisanactivatorofsmallconductanceKCa channelsthatdisplaysselectivityforSK2andSK3channels(EC50valuesare5.6and14µMforSK3andSK2respectivelyandefficacyis90%and71%forhSK3andhSK2,respectively)3.CyPPAdisplaysnoactivityatSK1andIK(KCa3.1)channels3.
CyPPAisatoolfordistinguishingSK2andSK3fromSK1andIK;nootheragentsdistinguishthesechannelsfromoneanother.Therefore,itenrichestherepertoireoftoolsavailabletotestthehypothesisthatSKchannelsmaybetargetsforfutureCNSdrugs4.
CyPPAwasfoundtoblockTRPM7channels,demonstratinganoverlapbetweenpharmacologicalcompoundsactingonTRPandKCachannels5.
CyPPA(#C-110) isahighlypure,synthetic,andbiologicallyactivecompound.
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多谢!
不知道能不能直接买到大鼠抗豚鼠血管IgG的ELISA试剂盒。
如果买不到这种试剂盒,用空白板的话(就是反应孔中没有抗原或者抗体包被),把自制的豚鼠血管抗原加到反应孔中进行孵育,能不能使抗原附着在孔壁上?这样的话就可以按照elisa的操作步骤进行大鼠血清检测了。
目前的设想是自制抗原,粉碎豚鼠的血管,制成悬液,通过反复多次的冻融,离心后收集上清液作为抗原(即豚鼠血管抗原)。
然后将抗原加入到elisa的反应孔中(这种是特制的空白板,就是反应孔中没有抗原或抗体包被)进行孵育,使豚鼠血管抗原附着在孔壁上,就是让反应孔充当固相载体,形成固相抗原。倒掉多余的抗原。
再加入待检测的大鼠血清,这样血清中的特异性抗豚鼠血管IgG就可以跟固相的抗原结合,形成固相抗原抗体复合物。
加入酶标的兔抗鼠或者羊抗鼠IgG,形成酶标的抗原抗体复合物。
然后就是一些显色步骤。
现在关键的问题就是抗原加入空白板,能不能形成固相抗原,如果不能跟孔壁附着的话,在后面洗涤的过程中就被洗掉了,那就没办法完成检测了。
请各位大侠给指条明路啊!!!小弟在此多谢了!!!
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