检测原理 在微孔板中存在竞争性测试化合物的情况下,将 GR 添加到荧光糖皮质激素配体 (Fluormone GS Red) 中。有效竞争剂的存在会阻止 GS Red/GR 复合物的形成,从而降低偏振度。测试化合物存在情况下的偏振度变化可用于测定测试化合物与 GR 的相对亲和力。该方法旨在通过使用辅激活因子相关稳定肽在室温下进行。或者,可以在不存在稳定肽的情况下在 4°C 下进行检测,此时 GR 结合特性的变化不明显。
如需大包装规格或帮助满足从检测方法开发到文库筛选等其他核受体检测需求,请联系我们。
仅供科研使用。不可用于诊断程序。
规格
适用于(应用)竞争性结合检测,核受体检测
配体GR
检测数量400 x 20 μL assays
产品类型检测试剂盒
数量400 x 20 μL assays
产品线PolarScreen™
Unit SizeEach
内容与储存
1管糖皮质激素受体重组蛋白(-68至 -85°C 储存) 1管 Fluormone GS Red 示踪剂(-5至 -30°C 储存) 3管 GR 筛选缓冲液(-5至 -30°C 储存) 3管 GR 稳定肽(-68至 -85°C 储存) 1管 DTT(-5至 -30°C 储存)
常见问题解答 (FAQ)
How do I export Ct values in the TaqMan hPSC Scorecard Analysis Software?
The only way to view the Ct values in the current version is through the expression plot. We expect the software's Excel export to include the gene names and Ct values in a future version of the software.
What are the common issues encountered with FP assays?
-No assay window: Check that your instrument is designed to do FP experiments.
-Poor discrimination between the minimum and maximum mP Control (both include the receptor): This may be an indication of the quality of the receptor. Do not make single use aliquots of the receptor or store diluted.
-The type of microplate is critical: Please see if the protocol specifies untreated polystyrene or NBS-coated plates. If the plate binds the fluorescent substrate, it cannot freely rotate and the mP reading will always be high. White plates cannot be used for FP assays. Avoid the use of silanized pipette tips and vials.
-The No Receptor Control' (free Fluormone/Tracer) should have much higher RFU values compared to buffer alone. If not, there is a problem with the instrument setup or less commonly with the Fluormone/Tracer.
-If the free Fluormone/Tracer mP value is greater than 100 mP or negative, please refer to the user manual for your instrument and reset the G factor.
What is the best type of microplate reader for fluorescence polarization (FP) assays?
For FP assays, plate readers must specifically have FP built in. Please see our instrument compatibility portal (http://www.thermofisher.com/us/en/home/industrial/pharma-biopharma/drug-discovery-development/dd-misc/instrument-compatibility-portal.html).
How can I test the fluorescence polarization (FP) settings of my microplate reader before doing the assay?
We offer a FP One-Step Reference Kit (https://www.thermofisher.com/order/catalog/product/P3088) to detect in green (Ex/Em = 488/535 nm) and red (Ex/Em = 525/590 nm) emission ranges, suitable for the PolarScreen Green and Red Assays. The alternative is to perform the Controls in the respective assay kit prior to performing the actual assay using test compounds.