LyophilizedPowderThisproductisfreezedried.Allwatermoleculeshavebeenremoved.
- PeptideNSSTEDGIKRIQDDC,correspondingtoaminoacidresidues4-18ofhumanAT1 receptor(Accession P30556).Extracellular,N-terminus.
CellsurfacedetectionofAT1 receptorinliveintacthumanTHP-1monocyticleukemiacells:___Cellsalone.
___Cells+RabbitIgGisotypecontrol-FITC.
___Cells+AngiotensinII ReceptorType-1(extracellular)-FITC Antibody(#AAR-011-F),(5µg).
CellsurfacedetectionofAT1 receptorinliveintacthumanJurkatT-cellleukemiacells:___Cellsalone.
___Cells+RabbitIgGisotypecontrol-FITC.
___Cells+AngiotensinII ReceptorType-1(extracellular)-FITCAntibody(#AAR-011-F),(5µg).
- 1.deGasparo,M.etal.(2000)Pharmacol.Rev.52,415.
- 2.Balakumar,P.andJagadeesh,G.(2014)J.Mol.Endocrinol.53,R71.
- 3.Graus-Nunes,F.etal.(2017)Mol.Cell.Endocrinol.439,54.
TheBIOLOGicalfunctionsof angiotensinII aremediatedby AT1 and AT2 receptors.BothbelongtotheG-proteincoupledreceptorsuperfamilyandarecomprisedofsevenhydrophobictransmembranesegmentsformingαhelicesinthelipidbilayerofthecellmembrane,anextracellularN-terminusandanintracellularN-terminus1.
Twohighlyhomologousisoformsofthereceptor,AT1A andAT1Bhavebeenidentifiedinrodents.TheAT1A receptoraccountsfor90%ofthetotalbinding,andispredominantinthekidney,vascularsmoothmusclecells,heart,liver,andinsomeareasofthebrain,whiletheAT1B receptorisfoundpredominantlyinthepituitaryandadrenalglands,placenta,lung,andbrain.Theseisoformsarepharmacologicallyindistinguishable andarebothselectivelyantagonizedby losartan.
ThroughAT1receptor,angiotensinIIstimulatesmultiplesignalingpathways,cross-talkswithseveraltyrosinekinases,andtrans-activatesgrowthfactorreceptors.PhosphorylationofthereceptorbyGPCRkinasesterminatesreceptoractivationandpromotesβ-arrestinrecruitment.β-arrestin-scaffoldedsignalingmediates‘secondarysignaling’thatinvolvesmultiplekinasesthatlinktocellprotectivedownstreamsignalingmolecules2.
Inamodelofdiet-inducedobesemice,AT1receptorinhibitiondemonstratesbeneficialpleiotropiceffects.Micereceivingthetreatmentexhibitenhancedpancreaticduodenalhomeobox1(PDX1)and GLP-1 pancreaticisletexpression.PDX1isletsareessentialfortheexpressionofglucokinaseand GLUT2.GreaterGLP-1levels,isletvascularizationandreducedapoptosisandmacrophageinfiltrationarealsoobserved3.
Anti-AngiotensinIIReceptorType-1(extracellular)Antibody(#AAR-011)isahighlyspecificantibodydirectedagainstanextracellularepitopeofthehumanprotein.Theantibodycanbeusedinwesternblot,immunohistochemistry,immunocytochemistry,andindirectflowcytometryapplications.IthasbeendesignedtorecognizeAT1 receptorfromrat,mouse,andhumansamples.
Anti-AngiotensinIIReceptorType-1(extracellular)-FITCAntibody(#AAR-011-F)isdirectlyconjugatedtofluoresceinisothiocyanate(FITC).Thislabeledantibodycanbeusedinimmunofluorescentapplicationssuchasdirectflowcytometryusinglivecells.
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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！！！
多谢！
