MarkerGeneTM Glucuronidase (GUS) Reporter GeneActivity Detection Kit
背景描述:
报告基因普遍用做“标志物 Markers”对基因调控和定位分析,以及突变基因分析。通过免疫实验,生化活性或组织化学分析来对组织或细胞的报告基因表达进行检测。
大肠杆菌来源的β-葡萄糖苷酶(β-glucuronidase,GUS)普遍认为是转基因植物研究中非常理想的报告基因(marker gene)。GUS报告基因系统具有许多优势,包括稳定表达大肠杆菌GUS,不会干扰正常植物代谢和高等植物内几乎无内源性GUS活性。
数种葡萄糖醛酸底物可用来检测GUS表达,这些底物都包含糖基D-葡萄糖醛酸苷(D-glucopyranosiduronic acid)通过糖苷键连接到生色底物、荧光底物或其他检测分子的—OH。体外用于GUS活性检测的最广泛使用的荧光底物为MUG(4-Methylumbelliferyl beta-D-glucuronide,4-甲基伞型酮-beta-D-葡糖苷酸,CAS:6160-80-1或881005-91-0)。一旦被水解,生成荧光素4-MU(4-甲基伞型酮)和β-D-葡萄糖醛酸。
检测原理:
本MarkerGeneTM GUS报告基因活性检测试剂盒(MarkerGeneTM Glucuronidase(GUS) Reporter Gene Activity Detection Kit)以MUG为荧光显色底物。当植物或其他细胞用GUS萃取液(含磷酸-EDTA,pH 7.0和去污剂)萃取后,所得到的萃取液(含GUS)能水解4-MUG生成4-MU(4-甲基伞型酮,pKa 8.2)和β-D-葡萄糖醛酸。加入碳酸钠缓冲液终止该反应(因4-MU在pH值高于pKa的情况下发射最大荧光信号)。4-MU的最大激发波长为365nm,最大发射波长为455nm。
试剂盒组分:
组分名称 |
组分编号 |
规格 |
保存 |
GUS Extraction Buffer |
0877-001 |
25 mL |
Store cold (4℃) |
Carbonate Stop Buffer |
0877-002 |
25 mL |
Store cold (4℃) |
GUS Assay Buffer |
0877-003 |
5 mL |
Store cold (4℃) |
4-MU calibration stock solution |
0877-004 |
5 mL |
Store cold (4℃) |
订购信息:
货号 |
产品名称 |
规格 |
价格(元) |
M0877 |
MarkerGene™ β-Glucuronidase (GUS) Reporter Gene Activity Detection Kit |
1Kit |
5680.00 |
使用文献(MarkerGene™ β-Glucuronidase(GUS) Reporter Gene Activity Detection Kit):
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