LyophilizedPowderThisproductisfreezedried.Allwatermoleculeshavebeenremoved.
AntigenIncl.ThisantibodyisshippedwithitsantigenFREEofcharge!
- PeptideCHSEDEKLSFEAVR,correspondingtoaminoacidresidues56-69ofhumanSTIM1(Accession Q13586). STIM1expressedontheplasmamembrane:Extracellular,N-terminus.STIM1expressedontheER:Luminal,N-terminus.
Expression ofSTIM1inratpancreasImmunohistochemicalstainingofratparaffin-embeddedpancreassectionsusingAnti-STIM1(extracellular)-ATTO-550 Antibody(#ACC-063-AO),(1:20),(red).A.STIM1stainingishighlyspecificforendocrinecellsintheIsleofLangerhans.B.Hoechst33342(blue)isusedasthecounterstain.C.MergedimageofpanelsAandB.
ExpressionofSTIM1inRBLcellsCellsurfacedetectionofSTIM1inliveRBLcells.A.ExtracellularstainingofcellswithAnti-STIM1(extracellular)-ATTO-550 Antibody(#ACC-063-AO),(1:20).B.Nuclearstainingofcellsusingthecell-permeabledyeHoechst33342.C.MergedimageofpanelsAandB.
- 1.Eisner,D.A.etal.(2005)Exp.Physiol.90,3.
- 2.Chakrabarti,R.andChakrabarti,R.(2006)J.Cell.Biochem.99,1503.
- 3.Feske,S.etal.(2006)Nature441,179.
- 4.Pickett,J.(2006)NatureReviewsMol.CellBiol.7,794.
- 5.Dziadek,M.A.andJohnstone,L.S.(2007)CellCalcium42,123.
- 6.Yuan,J.P.etal.(2007)Nat.CellBiol.9,636.
- 7.Muik,M.etal.(2008)J.Biol.Chem.283,8014.
- 8.Li,Z.etal(2007)J.Biol.Chem.282,29448.
- 9.Hewavitharana,T.etal.(2007)CellCalcium42,173.
CytosolicCa2+ haslongbeenknowntoactasakeysecondmessengerinmanyintracellularpathwaysincludingsynaptictransmission,musclecontraction,hormonalsecretion,andcellgrowthandproliferation.1,2 ThemechanismcontrollingtheinfluxofintracellularCa2+ eitherbycalcium-release-activatedCa2+ channels(CRAC)orfromintracellularstoreshaslatelybecomeofgreatinterest.
Recently,severalkeyplayersofthestore-operatedcomplexhavebeenidentified.3 TheOraifamilyconsistsofthreemembers,Orai1-3,andtheSTIMfamily,whichconsistsoftwomembers,STIM1andSTIM2.Orai1(alsoknownasCRACM1)actsasthestore-operatedcalciumchannel(SOC)andSTIM1astheendoplasmicreticulum(ER)Ca2+ sensor.3,4 ThemajorityofSTIM1appearstobelocalizedintracellularlyattheERmembranewhilelowexpressionofSTIM1hasbeendetectedonthecellsurfaceofseveralcelltypes.5 STIM1hasanamino-terminalEFhandCa2+ bindingdomainfacingthelumenoftheER.6 UponCa2+ storedepletion,STIM1moleculesareredistributedinpunctaeunderneaththeplasmamembraneandactivateSOCs.
SeveralpossIBLeinteractionsbetweenSTIM1andOrai1havebeensuggested.ThemostsimpleandcitedisadynamicinteractionbetweenthecytosolicC-terminusofSTIM1andthecytoplasmicdomainoftheOrai1channel.7-9 STIM1isassumedtoregulatetheactivityofallknownSOCs,includingnativeSOCs.5 ConsistentwiththeirimportantroleascalciumsensorswithintheER,STIM1proteinsareubiquitouslyexpressed.
Anti-STIM1(extracellular)Antibody(#ACC-063)isahighlyspecificantibodydirectedagainstanextracellularepitopeofthehumanStromalinteractionmolecule1.Theantibodycanbeusedinwesternblot,immunocytochemistry,immunohistochemistry,andindirectflowcytometryapplications.IthasbeendesignedtorecognizeSTIM1fromhuman,rat,andmousesamples.
Anti-STIM1(extracellular)-ATTO-550Antibody(#ACC-063-AO)isdirectlylabeledwithanATTO-550fluorescentdye.ATTOdyesarecharacterizedbystrongabsorption(highextinctioncoefficient),highfluorescencequantumyield,andhighphoto-stABIlity.TheATTO-550fluorescentlabelisrelatedtothewellknowndyeRhodamine6GandcanbeusedwithfilterstypicallyusedtodetectRhodamine.Anti-STIM1(extracellular)-ATTO-550AntibodyhasbeentestedinimmunocytochemistryandimmunohistochemistryapplicationsandisespeciallysuitedforexperimentsrequiringsimultaneouslabelingofdifferentMarkers.
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请问：同型对照应该怎么设？
不买专门的IgG2Bisotypecontrol，而用其他的抗体，
比如mouseanti-humanCD117，Igclass也是IgG2B，可以吗？
选用和特异性抗体同种亚型的，同样荧光素标记的抗体，采用同样的染色步骤染同样浓度的细胞。然后上机检测。如果细胞有非特异性的结合，那同型对照的荧光强度可能会比未染色的阴性对照要高一些。
同型对照只用于那种阳性和阴性之间没有明确分解的情况。如果阳性细胞群非常清楚，是不需要同型对照的。另外，在多色同染的情况下，也不采用同型对照，而是采用FMO（fluorensence minus one）对照。
另外，对于同型对照的使用，也有很多反对意见。就算是同型抗体，也可能会有不同的亲和力，并不一定就能真实反应非特异性的结合。
英 ['aɪsətaɪp]美 ['aɪsəˌtaɪp]
n. 同型动物（或植物），图形文字，象征性图像;同位型;同模;同号模式
同型对照（阴性对照）管：A同型+B同型+C同型
单阳性对照
A检测管：A+B同型+C同型
B检测管：B+A同型+C同型
C检测管：C+A同型+B同型
不存在荧光重叠的两个抗体不需要调节补偿。
样本为PBMC，想用三色标记Treg细胞，分别为CD4-PECY，CD25-Fitc，Foxp3-PE，想知道如何设置荧光补偿，用的是BD的流式仪，1、是设置阴性管一个，单阳性管每种一个，样品管一个么？
荧光素强度依次为PE， PECY5 APC FITC, PE最强，FITC最弱。 所以，建议抗体组合为：
Foxp3-PE，CD4-FITC，CD25-PECy5.5或者CD25-APC。
同时购买三个相应的同型对照抗体。
样品管设置如下：
1. cell + FITC-isotype + Pecy5.5（ or APC）-isotype + PE-isotype
2. Cell + FITC-isotype + CD25-mAb + PE-isotype
3. Cell + PE-mAb + FITC-isotype + Pecy5.5（ or APC）-isotype
4. Cell + CD4-FITC + Foxp3-PE + CD25 -PECy5.5( or APC)。 [这一管的数据可以拿来分析用]。
在充分了解调整电压和补偿原理的基础上，也可以做一些简化。如果不太懂，就按照这个来。其中Foxp3的抗体和同型对照抗体都是在固定破膜以后再加，不是与CD4或者CD25同时加。
