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Transheep推出雙熒光素酶報告基因系統(慢病毒介導)

 
        報告基因檢測,是真核基因表達調控研究的常用方法,常用于MirRNA靶點驗證,啟動子效率驗證,信號通路檢測等報告基因檢測相關實驗。由于檢測光量子的方法 非常敏感,采用生物發光(bioluminescent)法,是報告基因檢測最常用的有效手段。在用螢火蟲熒光素酶定量基因表達時 ,采用第二個報告基因來減少實驗的變化因素。 通常使用螢火蟲熒光素酶(Firefly luciferase)和海腎熒光素酶(Ranilla luciferase)催化的發光反應。雙熒光素酶報告基因測試系統在單管中進行雙熒光素酶報告基因測試,快速,靈敏,準確,簡便。






TsLVGLTM信號通路報告系統
TsLVGLTM信 號通路報告系統是權陽生物自主研發的用于檢測通路基因轉錄活性水平為目的的報告基因載體,含有若啟動子miniP,用于研究細胞內信號通路中轉錄應答元件 的活性。使用luc2為報告基因,表達效率遠高于pGL3系列的Luc+,優化后的luc具有更少的隱性調控序列,半衰期延長等優點。載體使用慢病毒骨架 系統,攜帶Puro抗性,可進行真核細胞系的篩選,實現穩轉。主要用于轉錄活性、腫瘤治療藥物的研發以及基因過表達和RNAi的表型分析等相關研究。



相關載體



預裝應答原件表

EnhancerElement Abbrv. Transcriptionfactor(s) Signaltransductionpathway(s)
Activatorprotein1 AP1 c-jun/c-fos JNK
cAMPresponseelement CRE ATF2/CREB ATF2/CREB
E-boxDNAbindingelement E-box Myc/Max cellproliferation
E2FDNAbindingelement E2F E2F/DP1 cellcycleprogression
Estrogenresponseelement ERE estrogenreceptor estrogenreceptor
Glucocorticoidresponseelement GRE GR glucocorticoid/HSP90
Heatshockresponseelement HSE HSF heatshockresponse
IFN-γactivationsequence GAS STAT1/STAT1 proliferation/inflammation
Interferon-stimulated ISRE STAT1/STAT2 proliferation/inflammation
responseelement
STAT3responseelementSTAT3 STAT3 STAT3/STAT3 proliferation/inflammation
NuclearfactorofactivatedT-cells NFAT NFAT PKC&Ca2+/calcineurin
NuclearfactorofκBcells NFκB NFκB NFκB
p53responseelement p53 p53 cellgrowth/apoptosis
Retinoicacidresponseelement RARE RAR retinoicacidreceptor
Rbresponseelement Rb Rb cellcycleprogression
Serumresponseelement SRE Elk-1/SRF MAPK/JNK
Thyroidresponseelement TRE Thyroidreceptor thyroidhormonereceptor

相關技術服務


相關載體

載體介紹 載體名稱 作用位點 出品公司
信號通路報告系統 pLvGL4-minipLuc2P   Transheep 
Rluc對照質粒 pLvGL4-minipLuc2P   Transheep 
Luc對照質粒 pTA-Luc   Clontech
信號通路報告系統 pGAS-TA-Luc STAT1/STAT1 Clontech
信號通路報告系統 pSTAT3-TA-Luc STAT3/STAT3 Clontech
信號通路報告系統 pISRE-TA-Luc STAT1/STAT2 Clontech
信號通路報告系統 pIκB-EGFP NFκB Clontech
信號通路報告系統 pNFAT-TA-Luc NFAT Clontech
信號通路報告系統 pCaspase3-sensor Caspase3-sensor Clontech
信號通路報告系統 pAP1(PMA)-TA-Luc c-jun/c-fos Clontech
信號通路報告系統 pCRE-Luc ATF2/CREB Clontech
信號通路報告系統 pGRE-Luc GR Clontech
信號通路報告系統 pHSE-Luc HSF Clontech
信號通路報告系統 pE2F-TA-Luc E2F/DP1 Clontech
信號通路報告系統 pTA-AP1-Luc c-jun/c-fos Promega
信號通路報告系統 pTA-CRE-Luc ATF2/CREB Promega
信號通路報告系統 pTA-E-box-Luc Myc/Max Promega
信號通路報告系統 pTA-E2F-Luc E2F/DP1 Promega
信號通路報告系統 pTA-ERE-Luc estrogenreceptor Promega
信號通路報告系統 pTA-GRE-Luc GR Promega
信號通路報告系統 pTA-HSE-Luc HSF Promega
信號通路報告系統 pTA-GAS-Luc STAT1/STAT1 Promega
信號通路報告系統 pTA-ISRE-Luc STAT1/STAT2 Promega
信號通路報告系統 pTA-STAT3-Luc STAT3/STAT3 Promega
信號通路報告系統 pTA-NFAT-Luc NFAT Promega
信號通路報告系統 pTA-NFκB-Luc NFκB Promega
信號通路報告系統 pTA-p53-Luc p53 Promega
信號通路報告系統 pTA-RARE-Luc RAR Promega
信號通路報告系統 pTA-Rb-Luc Rb Promega
信號通路報告系統 pTA-SRE-Luc Elk-1/SRF Promega
信號通路報告系統 pTA-TRE-Luc Thyroidreceptor Promega
信號通路報告系統 M50 Super 8x TOPFlash  TOPFlash    Transheep 
信號通路報告系統 pFA2-Elk1 FA2-Elk1 Stratagene
信號通路報告系統 pFC-MEKK FC-MEKK Stratagene
信號通路報告系統 pGL2Basic-EcadK1 EcadK1 Transheep 
信號通路報告系統 pGL2Basic-EcadK1/EpaIMUT/EboxMUT/Ebox2MUT EcadK1 Transheep 
信號通路報告系統 pGL3.0-IFNβ-luc   Transheep 
信號通路報告系統 pCRE-SEAP   Clontech
信號通路報告系統 TOPFlash   Millipore
信號通路報告系統 pGL4.10   Promega
信號通路報告系統 pGL4.11   Promega
信號通路報告系統 pGL4.13   Promega
信號通路報告系統 pGL4.15   Promega
信號通路報告系統 pGL4.17   Promega
信號通路報告系統 pGL4.19   Promega
信號通路報告系統 pGL4.22   Promega
信號通路報告系統 pGL4.26   Promega
信號通路報告系統 pGL4.27   Promega
信號通路報告系統 pGL4.29   Promega
信號通路報告系統 pGL4.30   Promega
信號通路報告系統 pGL4.75   Promega
信號通路報告系統 pGL4.79   Promega


相關試劑

      雙熒光素酶報告基因檢測試劑盒

  • 包裝清單:
產品編號 產品名稱 包裝
TsB0027-1 報告基因細胞裂解液 60ml×10
RG009-2 螢火蟲熒光素酶檢測試劑 10ml×10
RG009-3 Renilla熒光素酶檢測緩沖液 10ml×10
RG009-4 Renilla熒光素酶檢測底物 0.1ml×10
 
RG009-5 相關載體 5ug質粒&500ul菌液

  • 產品說明:
        Transheep生產的雙熒光素酶報告基因檢測試劑盒(Dual Luciferase Reporter Gene Assay Kit),是先以熒光素(luciferin)為底物來檢測螢火蟲熒光素酶(Firefly luciferase),后以coelenterazine為底物來檢測海腎熒光素酶(Renilla reniformis luciferase,簡稱Renilla luciferase),并且在后續加入Renilla luciferase的底物時同時加入了抑制Firefly luciferase催化luciferin發光的物質,使后續檢測僅僅測定到Renilla luciferase的活性,實現雙熒光素酶報告基因檢測。
        螢火蟲熒光素酶可以催化luciferin氧化成oxyluciferin,在luciferin氧化的過程中,會發出生物熒光 (bioluminescence)。海腎熒光素酶可以催化coelenterazine氧化成coelenteramide,在 coelenterazine氧化的過程中也會發出生物熒光。然后可以通過熒光測定儀也稱化學發光儀(luminometer)或液閃測定儀測定上述熒光 素酶催化的氧化過程中釋放的生物熒光。
        通過熒光素酶和其底物這一生物發光體系,可以極其靈敏、高效地檢測基因的表達。通常把感興趣
的 基因轉錄的調控元件克隆在firefly luciferase的上游或其他適當的地方,構建成報告基因質粒。然后轉染細胞,適當刺激或處理后裂解細胞,測定熒光素酶活性。通過熒光素酶活性的高低 判斷刺激前后或不同刺激對感興趣的調控元件的影響。Rinilla luciferase更多地被用作轉染效率的內參,以消除細胞數量和轉染效率的差異。
        螢火蟲熒光素酶催化luciferin發光的最強發光波長為560nm。海腎熒光素酶催化coelenterazine發
光的最強發光波長為465nm。
       本試劑盒可以測定1000個樣品。

  • 保存條件:
TsB0027-1和TsB0027-3在4℃保存1個月有效,-20℃保存一年有效,-70℃可以長期保存;TsB0027-2和
TsB0027-4在-20℃避光保存6個月有效,-70℃避光保存一年有效。

  • 注意事項:
        加入Renilla熒光素酶檢測工作液后對于上一步驟中的螢火蟲熒光素酶的抑制可以達到99%以上。但總會殘留微量活性,因此,宜在轉染時把 Renilla熒光素酶的表達量控制在其RLU讀數高于螢火蟲熒光素酶RLU讀數的10%。Renilla熒光素酶的讀數高于螢火蟲熒光素酶的讀數是完全 可以的,通常不會有明顯的負面影響。
        為取得最佳測定效果,在用單管的熒光測定儀測定時,樣品和測定試劑混合后到測定前的時間應盡
量控制在相同的時間內,例如30秒內。
        由于溫度對酶反應有影響,所以測定時樣品和試劑均需達到室溫后再進行測定。
        為保證熒光素酶檢測試劑的穩定性,可以采取適當分裝后避光保存的方法,以避免反復凍融和長時
間暴露于室溫。一般來說,如果不分裝使用三次(期間凍融三次),對測定結果無明顯影響。
        樣品和測定試劑混合后,必須等待1-2秒,再進行測定。測定時間通常為10秒,根據情況也可以測定更長或更短時間,但是同一批樣品最好使用相同的測定時間。

  • 特別注意:Renilla熒光素酶檢測工作液后需配制后立即使用,不可配制成工作液后長期保存。
Renilla熒光素酶檢測工作液不太穩定,配制后如果不能立即使用在-20℃可以保存5-10天左右活力會有輕微下降,室溫放置一天會導致活力下降5-10%或更多,4℃放置10天左右會導致活力下降10-20%或更多。
        TsB0027-4 Renilla熒光素酶檢測底物(100X)配制在無水乙醇中。由于無水乙醇容易揮發,有時會在初次使用時發現體積明顯小于0.1ml的情況,此時用無水乙醇把體積補足至0.1ml,并混勻后即可使用。
       為了您的安全和健康,請穿實驗服并戴一次性手套操作。
云南11选5预测
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