| productinformation
| Background | | STN7(serine/threonineproteinkinase)(EC=2.7.11.1)inArABIdopsisthaliana,anditsStt7homologueinChlamydomonasreinhardtii,isrequiredforLHCIIphosphorylationandforstatetransitions.Thelossofthisthylakoid-associatedkinaseblockedstn7(stt7)-deficientmutantsinstate1.Alternativenames:ProteinSTATETRANSITION7,Stt7homolog. | Immunogen | | KLH-conjugatedsyntheticpeptide(aminoacids128–141)specificforArabidopsisthalianaSTN7serine/threoninekinaseAt1g68830(Q9S713) | Host | | Rabbit | Clonality | | Polyclonal | Clone | | | Purity | | AffinitypurifiedseruminPBSpH7.4. | Format | | Lyophilized | Quantity | | 50µg | Reconstitution | | Forreconstitutionpleaseadd50µlofsterilewater. | Storage | | Storelyophilized/reconstitutedat-20°C;oncereconstitutedmakealiquotstoavoidrepeatedfreeze-thawcycles.Please,remembertospintubesbrieflypriortoopeningthemtoavoidanylossesthatmightoccurfromlyophilizedmaterialadheringtothecaporsidesofthetubes. | Testedapplications | | Westernblot(WB) | Relatedproducts | | AS101601|anti-STN8kinase|serine/threonine-proteinkinaseSTN8,chloroplasticforArabidopsisthaliana,rabbitantibodies
AS153080|anti-STN7|serine/threonine-proteinkinaseSTN7,chloroplasticforChlamydomonasreinhardtii,rabbitantibodies Secondaryantibodies | Additionalinformation | | AnextractfromSTN7mutantneedstobeusedinparareltodeterminespecificbandofSTN7proteinonawesternblot. |
applicationinformation
| Recommendeddilution | | 1:2000(WB) | Expected|apparentMW | | 63.2|44kDa(onureagel),55kDa(nourea) | Confirmedreactivity | | Arabidopsisthaliana,Nicotianatabacum | Predictedreactivity | | Glycinemax,Oryzasativa,Ricinuscommunis,Zeamays,Vitisvinifera
| Notreactivein | | Chlamydomonasreinhardtii | Additionalinformation | | Forbestresultswiththisantibodysamplebufferneedstocontain6Murea(138mMTrisHClpH6.8,6Murea,22.2%Glycerol,4.3%SDS). STN7isanuclearencodedproteinwhichislocalizedinchloroplast,hencepossesachloroplastictargetpeptides(cTP)atthebeginningoftheaminoacidsequencewhichiscutoff.AccrodingtoTargetP(programthatpredictsthelengthofthecTP:s)thelengthofcTPforStn7is45aa.STN7porteinmobilitycanbealsoaffectedbyureapresentinthegel. | Selectedreferences | | Fristedtetal.(2017).PSB33sustainsphotosystemIID1proteinunderfluctuatinglightconditions.JournalofExperimentalBotanydoi:10.1093/jxb/erx218. Mekalaetal.(2015).Plantsactivelyavoidstate-transitionsuponchangesinlightintensity-roleoflight-harvestingcomplexIIproteindephosphorylationinhighlight.PlantPhysiol.2015Apr22.pii:pp.00488.2015. Floodetal.(2014).NaturalvariationinphosphorylationofphotosystemIIproteinsinArabidopsisthaliana:isitcausedbygeneticvariationintheSTNkinases?PhilosTransRSocLondBBiolSci.2014Mar3;369(1640):20130499.doi:10.1098/rstb.2013.0499.Print2014. Yinetal.(2012).PhotosystemIIFunctionandDynamicsinThreeWidelyUsedArabidopsisthalianaAccessions.PLOSONE,openaccess. |
applicationexample 
Thylakoids3ugofchlorophyll/lanefromArabidopsisthalianawereextractedaccordingtoSirpiöetal(2011,MethodsMolBiol.2011;775:19-30)andwereseparatedon12%SDS-PAGEcontaining6Mureaandblotted1htoPVDF.Blotswereblockedwith5%milkinTTBSfor1hatroomtemperature(RT)withslowagitation.Blotwasincubatedintheprimaryantibodyatadilutionof1:1000overnightat4°Cwithslowagitationin1%milkTTBS.Theantibodysolutionwasdecantedandtheblotwasrinsedbrieflyonlyonce,thenwashedoncefor15minonlyinTBS-TatRTwithvigorousagitation.Blotwasincubatedinsecondaryantibody(anti-rabbitIgGhorserADIshperoxidaseconjugated)dilutedto1:10000in1%milkinTTBSfor2hat4°Cwithslowagitation.Theblotwaswashedasaboveandincubatedfor5minwithECL-solutionaccordingtothemanufacturersinstructions.Exposuretimewas1min. Toavoidcross-reactivitywithLHCproteinsagelcanberunfurthertoletproteinsbelow25kDaleaveagel. CourtesyDr.MaijaHolmström,UniversityofTurku,Finland Agrisera肽合成选择最合适的蛋白质序列以产生抗肽血清是成功免疫计划的第一步,也是最关键的一步。理想的肽序列应该是目标蛋白所特有的,并位于表位与抗体相互作用的最佳位置,因为某些抗肽抗体在某些应用中用于靶向天然蛋白时可能无法识别其表位。此外,必须考虑技术要求(例如,合成的适用性,结合以及免疫的预期溶解度)。Agrisera提供通过固相肽合成(使用Fmoc-Chemistry)生产的定制合成肽,包括末端半胱氨酸,并可能进行可选修饰(例如,乙酰化,生物素化和荧光团偶联)。肽可以从免疫级纯度(> 70%,足以生成或测试抗体)合成到更高的纯度(> 95%,在酶学和生物活性研究等方面需要)。纯度通过HPLC和质谱分析(根据需要进行氨基酸分析和测序)控制。我们的肽服务可以单独使用,也可以与我们的抗体生产服务结合使用。标准设计和合成服务根据您提交的目标蛋白质序列(提供为.txt文件或登录号提供),我们建议最多3个肽段序列被认为适合合成。所选择的序列(最多14个氨基酸+末端半胱氨酸)以15 mg免疫级纯度等级合成,没有任何进一步的修饰。交货时间为4周。重要说明:我们的标准服务不包含对目标生物体内与非靶标蛋白质的预期交叉反应的详细分析。但是,将考虑提供有关应排除交叉反应的蛋白质序列(.txt文件或登录号)的信息。如果可能,请在您希望优先(或避免)靶向区域,信号肽或任何已知的靶蛋白修饰(例如磷酸化位点)上指定其他信息。与载体蛋白的偶联由于与宿主动物的降解减少,通过与更高质量的载体蛋白偶联,合成肽的抗原性大大提高。与标准肽合成结合,可以将一部分肽(通常为2 mg)与以下标准载体蛋白之一偶联:KLH-匙孔戚血蓝蛋白是一种节肢动物和软体动物中的含铜蛋白。其分子大小(> 450 kDa)随其解离状态而变化,具体取决于pH。Agrisera提供了多克隆抗KLH抗体。BSA -67 kDa的牛血清白蛋白亚基是一种蛋白质,常用于免疫测定中,以阻断非特异性结合位点。如果将来的分析涉及BSA(例如,用作封闭剂),则不应将其用作载体。Agrisera提供了多克隆抗BSA抗体。有关我们标准肽服务可能进行的定制改编的其他信息(例如,更多的肽,或需要复杂肽设计的个别帮助),请通过info@agrisera.com与我们联系。
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