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GLuc-ON™ 启动子报告克隆

产品简介

GLuc-ON™ 启动子报告克隆在GLuc报告基因上游插入一段1.2~1.5 kb的序列,这段插入序列与基因转录起始位点(TSS)上游大约1.5kb到下游200bp之间的5'侧翼序列一致,可以通过观察荧光素酶的活性来研究启动子对基因的调节作用。

GeneCopoeia可提供预制和定制的GLuc-ON™ 启动子克隆,包括单报告基因载体系统和双报告基因载体系统。单报告基因载体系统以Gluc、 mCherry或GFP作为启动子的报告基因;双报告基因载体系统以Gluc作为启动子报告基因,以分泌型碱性磷酸酶 (SEAP)作为信号标准化的内参,可在活细胞中对超过20,000种人和18,000种小鼠启动子进行实时活性分析。

Promoter reporter clones mechanism of action

图1. GLuc-ON™ 启动子报告克隆的双报告基因检测原理图。

 

优势

活细胞分析

  • 分泌型Gluc报告子
  • 无需裂解细胞
  • 节省样本,减少突变,简化试验流程(如脉冲追踪分析等)

实时研究

  • 可获得即时检测数据
  • 与实时活性分析过程类似

分泌型双报告系统

  • 分泌型Gluc和分泌型碱性磷酸酶
  • 对多样本常规转染进行精确比较

高通量兼容

  • 可用于信号通路研究
  • 可进行高通量研究

高灵敏度

  • GLuc比萤火虫或Renilla荧光素酶灵敏度高1000倍

使用方便

  • 所有启动子报告克隆均可直接用于细胞转染

 

Gaussia 荧光素酶

GLuc-ON™ 启动子克隆采用经过修饰的GLuc(mGLuc)作为报告基因,能产生强烈且稳定的荧光信号,克服了人野生型GLuc(wtGLuc)信号衰退快的缺点。

Signal stability of mGluc and wtGlucSignal stability of mGluc and wtGluc

图2. mGLuc (蓝色) 和 wtGLuc (红色)的信号稳定性比较。左侧:含有稳定剂的缓冲液;右侧:常规缓冲液。

 

双报告系统

GLuc-ON™ 启动子克隆可以将分泌型碱性磷酸酶(SEAP)作为第二个报告子和内参。这种双报告系统可以对多样本常规转染进行精确比较。

 

图3. 不同启动子在H1B1B和HEK293T细胞中的活性分析。双报告系统启动子克隆和对照以两个重复转染进两种细胞中。转染24小时((HEK293T)和48小时(H1B1B)后分析样品。NEG(包含非启动子序列)和EMPTY(载体不含启动子)作为阴性对照。

载体和对照

载体类型

载体 报告基因 内参基因 筛选标记 载体类型
pEZX-PG04 Gaussia luciferase (GLuc) 分泌型碱性磷酸酶(SEAP) Puromycin 非病毒
pEZX-PG02 Gaussia luciferase (GLuc) N/A* Puromycin 非病毒
pEZX-PF02 eGFP N/A* Puromycin 非病毒
pEZX-PM02 mCherry N/A* Puromycin 非病毒
pEZX-LvPG04 Gaussia luciferase (GLuc) 分泌型碱性磷酸酶((SEAP) Puromycin 慢病毒
pEZX-LvPG02 Gaussia luciferase (GLuc) N/A* Puromycin 慢病毒
pEZX-LvPF02 eGFP N/A* Puromycin 慢病毒
pEZX-LvPM02 mCherry N/A* Puromycin 慢病毒
pEZX-LvPM03 tdTomato N/A* Puromycin 慢病毒

* SEAP可以在分泌型载体中表达。

 

对照克隆和SEAP表达克隆

购买 货号 产品 描述 载体 价格(¥)
GAPDH-PF02 GAPDH positive control clone House keeping gene promoter positive control in single GFP reporter vector GAPDH positive control clone 875
GAPDH-LvPF02 GAPDH positive control clone House keeping gene promoter positive control in single GFP reporter lentiviral vector GAPDH positive control clone 875
GAPDH-PG02 GAPDH positive control clone House keeping gene promoter positive control in single-reporter vector GAPDH positive control clone 875
GAPDH-LvPG02 GAPDH positive control clone House keeping gene promoter positive control in single-reporter lentiviral vector GAPDH positive control clone 875
GAPDH-PG04 GAPDH positive control clone House keeping gene promoter positive control in dual-reporter vector GAPDH positive control clone 875
GAPDH-LvPG04 GAPDH positive control clone House keeping gene promoter positive control in dual-reporter lentiviral vector GAPDH positive control clone 875
GAPDH-PM02 GAPDH positive control clone House keeping gene promoter positive control in single mCherry reporter vector GAPDH positive control clone 875
GAPDH-LvPM02 GAPDH positive control clone House keeping gene promoter positive control in single mCherry reporter lentiviral vector GAPDH positive control clone 875
NEG-PF02 Negative control clone Non-promoter sequence negative control in single GFP reporter vector Negative control clone 875
NEG-LvPF02 Negative control clone Non-promoter sequence negative control in single GFP reporter lentiviral vector Negative control clone 875
TR001 TRE negative control clone pEZX-PG02 2400
NEG-LvPG02 Negative control clone Non-promoter sequence negative control in single-reporter lentiviral vector Negative control clone 875
NEG-PG04 Negative control clone Non-promoter sequence negative control in dual-reporter vector Negative control clone 875
NEG-LvPG04 Negative control clone Non-promoter sequence negative control in dual-reporter lentiviral vector Negative control clone 875
NEG-PM02 Negative control clone Non-promoter sequence negative control in single mCherry reporter vector Negative control clone 875
NEG-LvPM02 Negative control clone Non-promoter sequence negative control in single mCherry reporter lentiviral vector Negative control clone 875
SEAP-PA01 Secreted alkaline phosphatase (SEAP)
Expression clone
Express SEAP.
Can be used for co-transfection with promoter clone
pEZX-PA01 875
SEAP-LvPA02 Secreted alkaline phosphatase (SEAP) expression clone Express SEAP in a lentiviral vector. Can be used for co-transduction with a promoter clone pEZX-PA02 875

 

Tear Away License

Tear away license for gaussia luciferase users

订购

预制的启动子报告克隆

GeneCopoeia提供超过20,000种人和18,000种小鼠不同载体的预制启动子报告克隆。

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慢病毒载体的启动子报告克隆

GeneCopoeia提供客户定制的慢病毒载体启动子报告克隆。 了解慢病毒载体价格信息,请查询

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定制启动子报告克隆

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需要定制克隆来比较启动子不同长度或不同区域的功能,请发邮件到 support@igenebio.com 或拨打4006-020-200

 

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