

Thisproductisfreezedried.Allwatermoleculeshavebeenremoved.
- Peptide(C)DDYGRPGIEKFREE,correspondingtoaminoacidresidues216-229ofhumanCaSR(Accession P41180). Extracellular,N-terminus.
- CellsurfacedetectionofCaSRinliveintacthumanTHP-1monocyticleukemiacells:___ Cellsalone.
___ Cells+RabbitIgGisotypecontrol-FITC.
___ Cells+ Anti-CalciumSensingReceptor (extracellular)-FITCAntibody(#ACR-004-F),2.5µg.CellsurfacedetectionofCaSRinliveintactmouseJ774macrophagecells:___ Cellsalone.
___ Cells+RabbitIgGisotypecontrol-FITC.
___ Cells+ Anti-CalciumSensingReceptor (extracellular)-FITCAntibody(#ACR-004-F),5µg.
- 1.Riccardi,D.andBrown,E.M.(2010)Am.J.Physiol.298,F485.
- 2.Hendy,G.N.etal.(2013)BestPract.Res.Clin.Endocrinol.Metab.27,285.
Thecalcium-sensingreceptor(CaSR)isaclassCG-protein-coupledreceptorencodedbyasinglecopygene.CaSRplaysamajorroleinthemaintenanceofaphysiologicalserumionizedCa2+ concentrationbyregulatingthecirculatinglevelsofparathyroidhormone1.
TheN-terminusofthereceptoriscomprisedofasignalpeptideof19 aminoacids.ThisterminusdirectsthenascentCaSRpolypeptideintothelumenoftheendoplasmicreticulum(ER)andisthencleaved.Theextracellulardomain(ECD)ofthematureproteinhasapproximately600 aminoacids,comprisedofabi-lobedVenus-flytrap-likedomain(VFT)connectedbyacysteine-richregionandashortpeptidelinkertotheseventransmembranedomain(TMD)withanintracellularC-terminaltailof216 aminoacids.CaSRfunctionsasahomodimerwithbothcovalentandnoncovalentinteractionsbetweentheVFTdomainsofeachmonomer. Ca2+ bindsinthecleftbetweenthetwolobesofeachVFTcausingthelobestocloseononeanotherandtheVFTtorotatewithtransferofaconformationalchangetothecysteine-richregion.
Intheparathyroidgland,up-regulationofCaSRby1,25-dihydroxyvitaminD[1,25(OH)2D]makestheglandmoreresponsivetoextracellularCa2+ andparathyroidhormone(PTH)secretionisreduced.Whilesomaticmutationsin CASR arerare,parathyroidglandsfrompatientswithprimaryorsevereuremicsecondaryhyperparathyroidismhavereducedCaSRexpression2.
Anti-CalciumSensingReceptor(extracellular)Antibody(#ACR-004)isahighlyspecificantibodydirectedagainstanepitopeofthehumanCaSR.Theantibodycanbeusedinwesternblot,immunohistochemistry,indirectflowcytometry,andimmunocytochemistryapplications.IthasbeendesignedtorecognizeCaSRfromhuman,rat,andmousesamples.
Anti-CalciumSensingReceptor(extracellular)-FITCAntibody(#ACR-004-F)isdirectlyconjugatedtofluoresceinisothiocyanate(FITC).Thislabeledantibodycanbeusedinimmunofluorescentapplicationssuchasdirectflowcytometryusinglivecells.
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此酶溶于水和50%饱和度以下的硫酸铵溶液。
(2)抗体如何选择?——根据目标分子的标签类型,检测的方法、抗体的适用范围、商家有没有提供自身质检结果等进行选择。
想做个蛋白的定位,可是没有好抗体,就想做个转基因小鼠,在基因后面加一个标签。但是之前没有相关经验,不知道哪一种tag更好呢,更适合在小鼠里表达呢?因为我蛋白比较小,加GFP那么大的话可能会影响表达。比较小的tag例如HA,Flag还有His等,不知道哪个更适合一些?求大神们给些经验。
单抗标签选择的一抗范围比较小一些,多抗标签选择的一抗范围可能相对大一些。而商品化二抗基本没什么差别。不管是选择什么样的抗体,在Western blot检测方面都是一样的
所以做Western blot的单抗和多抗标签并没有什么不同
求助:为什么用蛋白抗体仅出来一条带,而用Flag抗体则出来两条带?
谢谢大家啊
图一:蛋白抗体
图二Flag抗体
(2)6 X His tag 序列:CAT CAT CAC CAT CAC CAT
(3)GST标签序列:
1ATGTCCCCTATACTAGGTTATTGGAAAATTAAGGGCCTTGTGC
301 AACCCACTCGACTTCTTTTGGAATATCTTGAAGAAAAATATGAAGAGCAT
351 TTGTATGAGCGCGATGAAGGTGATAAATGGCGAAACAAAAAGTTTGAATT
401 GGGTTTGGAGTTTCCCAATCTTCCTTATTATATTGATGGTGATGTTAAAT
451 TAACACAGTCTATGGCCATCATACGTTATATAGCTGACAAGCACAACATG
501 TTGGGTGGTTGTCCAAAAGAGCGTGCAGAGATTTCAATGCTTGAAGGAGC
551 GGTTTTGGATATTAGATACGGTGTTTCGAGAATTGCATATAGTAAAGACT
601 TTGAAACTCTCAAAGTTGATTTTCTTAGCAAGCTACCTGAAATGCTGAAA
651 ATGTTCGAAGATCGTTTATGTCATAAAACATATTTAAATGGTGATCATGT
701 AACCCATCCTGACTTCATGTTGTATGACGCTCTTGATGTTGTTTTATACA
751 TGGACCCAATGTGCCTGGATGCGTTCCCAAAATTAGTTTGTTTTAAAAAA
801 CGTATTGAAGCTATCCCACAAATTGATAAGTACTTGAAATCCAGCAAGTA
851 TATAGCATGGCCTTTGCAGGGCTGGCAAGCCACGTTTGGTGGTGGCGACC
901 ATCCTCCAAAATCGGATCTGGTTCCGCGTGGATCCCCGGAATTCCCGGGT
951 CGACTCGAGCGGCCGCATCGTGACTGA

