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UniOne® TR-FRET Human CRBN Ligand Screening Assay Kit
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对比
咨询Component | Concentration | Volume(100T) | Volume(500T) | Volume(2500T) | Volume(10000T) | Storage Condition |
|---|---|---|---|---|---|---|
Thalidomide | 10 mM in DMSO | 5 μL | 5 μL | 25 μL | 100 μL | -80℃ |
Tag1-CRBN/DDB1 | 100× | 5μL | 25μL | 125μL | 500 μL | -80℃ |
Thalidomide-Ac | 100× | 5μL | 25μL | 125μL | 500 μL | -80℃ |
Anti-Tag1-Eu | 50× | 10μL | 50μL | 250 μL | 1000 μL | -80℃ |
Detection Buffer | 10× | 250μL | 1.2mL | 6mL | 21 mL | -80℃ |
*Reagents should be protected from repeated freeze-thaw cycles. It is recommended to aliquot after the first thaw and store according to the storage conditions. The shelf life is one year.
*Thaw reagents on ice before use and equilibrate to room temperature.
*Prepare all reagent working solutions freshly before use.
人Cereblon (CRBN) 蛋白配体结合检测试剂盒采用均相时间分辨荧光技术（TR-FRET），可用于CRBN抑制剂或配体的结合检测和筛选，是实现快速检测CRBN抑制剂或配体的高通量筛选形式，基本原理如下图所示。
Eu标记的检测试剂（Anti-Tag1-Eu）特异识别结合Tag1-CRBN/DDB1蛋白，受体标记的沙利度胺（Thalidomide-Ac）与CRBN结合，使Eu和Ac靠近，在外部光源的激发下，供体与受体之间发生能量共振转移，检测665nm波长的信号强度即可测定CRBN/DDB1与Thalidomide-Ac的结合。反应体系中加入的CRBN配体或PROTAC竞争结合CRBN/DDB1，使信号降低，配体或PROTAC浓度升高，信号降低。该均相实验操作简单，无需洗涤步骤。
-80°C