Secrete-Pair™ Dual Luminescence Assay Kit
Secrete-Pair™ Dual Luminescence Assay Kit用于检测双报告系统中Gaussia Luciferase (GLuc)和Secreted Alkaline Phosphatase (SEAP)的活性。GLuc和SEAP都是分泌型蛋白,无需裂解细胞就能从培养基中获得实验样本。 |
试剂盒提供两种用于Gluc分析的缓冲液。Buffer GL-S含有稳定剂,弥补了Gluc信号快速衰退的缺点,可用于稳定活性分析。Buffer GL-H适用于检测低表达量的Gluc活性,可以提供更高的灵敏度。
试剂盒能检测双报告信号并能对转染标准化。标准化的Gluc活性可通过与未转染的空白样本比较获得。
Secrete Pair™ Gaussia Luciferase Assay Kit
Secrete Pair Gaussia Luciferase Assay Kit只能用于检测Gaussia luciferase。
Secrete Pair kits可与 GLuc-ON™启动子报告克隆和miTarget™ miRNA 3′ UTR克隆配套使用。
产品优势
活细胞分析
- 分泌型GLuc和SEAP
- 无需裂解细胞
实验条件更灵活
- 根据不同情况提供两种用于Gluc活性分析的缓冲液
- 使用稳定活性缓冲液可以在实验开始10分钟内保证90%以上的信号,并能延长发射光的半衰期30分钟以上
- 灵敏度高的缓冲液可以检测低表达量的GLuc
实时研究
- 快速获得实验数据
- 与实时活性研究类似
双报告系统检测
- 检测GLuc和SEAP
- 实现转染标准化,可用于样本间的比较
高通量兼容
- 快速简便的分析模式
- 可检测大量样品
载体选择
GeneCopoeia 提供Gaussia luciferase(Gluc)报告载体,可供广大研究者自由操作;只需将目的片段插入报告载体,即可应用荧光素酶检测试剂。 点击载体名称可查看载体图谱信息。
货号 | 载体 | 启动子 | 报告基因 | 示踪基因 | 筛选标记 | 抗性 | 载体类型 |
ZX101 | pEZX-GN01 | N/A* | Gluc | N/A* | Puromycin | Kan | Non-viral |
ZX102 | pEZX-GN03 | miniCMV | Gluc | N/A* | Puromycin | Kan | Non-viral |
ZX103 | pEZX-GA01 | N/A* | Gluc | Secreted alkaline phosphatase (SEAP) | Puromycin | Kan | Non-viral |
ZX104 | pEZX-GA02 | SV40 | Gluc | Secreted alkaline phosphatase (SEAP) | Neomycin | Amp | Non-viral |
ZX105 | pEZX-GA03 | miniCMV | Gluc | Secreted alkaline phosphatase (SEAP) | Puromycin | Kan | Non-viral |
ZX106 | pEZX-LvGN01 | N/A* | Gluc | N/A* | Puromycin | Amp | Lentiviral |
ZX107 | pEZX-LvGA01 | N/A* | Gluc | Secreted alkaline phosphatase (SEAP) | Puromycin | Amp | Lentiviral |
* A separate vector is available for SEAP expression.
操作流程
Figure 1.不同缓冲体系中GLuc的信号稳定性比较。细胞转染人类Gluc报告基因克隆后收集细胞培养物,每个反应10μl。根据试剂盒的实验步骤进行操作,对每个试剂盒中两种缓冲体系进行测试,获得的信号百分比(Y轴)作为信号稳定性的指标。实验结果如图所示,在两种试剂盒中(Secrete-Pair和竞争者Gaussia luciferase assay kit),含有稳定剂(-S)的缓冲体系中Gluc的活性比不含稳定剂(-H)的缓冲体系中稳定。不仅如此,Secrete-Pair试剂盒缓冲体系中Gluc信号比竞争者试剂盒中稳定性更高(实验开始10分钟内近90%的信号可被捕获)。
Figure 2. GLuc和SEAP分析。细胞转染GLuc-SEAP双报告基因克隆后收集细胞培养物,每个反应10μl。反应初期,GL-H缓冲液中wtGLuc活性比GL-S缓冲液中高3-5倍。但GL-S缓冲液中GLuc活性更稳定。
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