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細(xì)胞應(yīng)激傳感器是一種基因編碼的熒光生物傳感器,當(dāng)細(xì)胞承受內(nèi)質(zhì)網(wǎng)(ER介導(dǎo)的)應(yīng)激或經(jīng)歷未折疊蛋白反應(yīng)(UPR)時,它會產(chǎn)生非常亮的綠色熒光。重要的是,綠色熒光信號是核定位的,用于簡單的細(xì)胞分割和圖像分析。該傳感器基于在內(nèi)質(zhì)網(wǎng)應(yīng)激期間由IRE1蛋白剪接的XBP1 mRNA[Iwawaki等人2003]。
該產(chǎn)品用于測量96孔板活細(xì)胞上的細(xì)胞應(yīng)激反應(yīng)。它已在活體HEK293T細(xì)胞以及iPSC衍生的心肌細(xì)胞和神經(jīng)元中得到驗證。可以在活細(xì)胞成像系統(tǒng)、自動熒光板閱讀器或熒光顯微鏡上檢測到測定的熒光。
艾美捷 Montana Molecular綠細(xì)胞應(yīng)激測定試劑盒#U0900G包含的材料:
綠色細(xì)胞應(yīng)激感應(yīng)器BacMam
丁酸鈉,500 mM水溶液
芍藥苷100 μM(對照品)
存儲:BacMam存貨應(yīng)在4℃的溫度下儲存,并在原始包裝中加以保護(hù)。儲存HDAC內(nèi)部溫度為4℃。儲存溫度控制在-20℃。避免重復(fù)凍融循環(huán)。我們建議在儲存超過12個月后重新測試BacMam庫存。如果您的BacMamstock已被純化,請在30天內(nèi)使用以獲得結(jié)果

HEK293T表達(dá)細(xì)胞應(yīng)激傳感器和野生型(WT)或野生型視紫紅質(zhì)或超氧化物歧化酶(SOD)的細(xì)胞。每種疾病突變蛋白都是一種已知的突變,導(dǎo)致蛋白質(zhì)錯誤折疊或聚集。紅細(xì)胞色素P23H導(dǎo)致視網(wǎng)膜色素變性,SOD G85R導(dǎo)致ALS。表達(dá)18小時后對細(xì)胞進(jìn)行分析。
Montana Molecular綠細(xì)胞應(yīng)激測定試劑盒文獻(xiàn)參考:
1.Iwawaki,Takao,Ryoko Akal,Kenji Kohno,and Masayukl Miura.2003."ATransgenlc Mouse Model for Monitor-ing Endoplasmic Reticulum Stress"NatureMedicine 10(1).NaturePublishing Group:98-102.
2.Rong,Juan,lanPass,PaulW.Dlaz,Tram A.Ngo,Michelle Sauer,GavinMagnuson,Fu-Yue Zeng,etal.2015."CellBasedHigh-Throughput Luciferase ReporterGene Assaysforldentifying and ProfilingChemicalModulatorsofEndoplasmic ReticulumSignaling Protein,IRE1.Journal of Biomolecular Screening 20(10).SAGEPublications:1232-45.
3.GrahamFL,Smiley J,Russell WC,Nairn R:CharacterlstIcsof a human cell lIne transformed byDNAfromhuman adenovlrus type5.JGen VIrol 1977,36(1):59-74.
4.DulbeccoR and Vogt M:Plaque formatlonand Isolatlon ofpure llneswith pollomyelltlsviruses.The Jour-nalof experimental medicine 1954.
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6.KostT,Condreay J,Ames R,Rees S,Romanos M:ImplementatlonofBacMam vIrus gene dellvery technol-ogy In adrug discovery settIng.Drug DiscoveryToday 2007,12(9-10):396-403
7.Tewson PH,Martinka S,Shaner N,HughesTE,Quinn AM:NewDAG andcAMP sensors optimized for live cellassaysin automatedlaboratoriles.Journal of BlomolecularScreening 2015.
8.Shaner,N.C,Lambert,G.G,Chammas,A,Ni,Y,Cranfil,PJ,Baird,M.A.,Sell,B.R,Allen,JR,Day,R.N,Israelsson M,Davidson,M.W,&Wang,J.(2013)"A bright monomericgreen fluorescent protein derivedfrom Branchios-toma lanceolatum."Nature Methods,May;10(5):407-9.dot:10.1038/nmeth.2413.
Montana Molecular產(chǎn)品主要包括GPCR檢測、細(xì)胞應(yīng)激和毒性相關(guān)檢測工具,為客戶提供了長時間內(nèi)觀察候選藥物對細(xì)胞過程影響的能力。艾美捷科技是Montana Molecular的中國代理商,為科研工作者提供優(yōu)質(zhì)的產(chǎn)品與服務(wù)。
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