The MST measurement for the binding of compounds to ASIC1a was performed using Monolith NT.115 (NanoTemper). In brief, the HEK293T cell was collected after 48 h transfection with eGFP-tagged wild-type or mutant ASIC1a plasmid and lysed by M-PER mammalian protein extraction reagent (Thermo ...
With the use of conventional capillaries and Monolith NT.115 (NanoTemper Technologies GmbH, Germany), MST was used to ascertain the binding affinity of GJB2 with Salvianolic acid B. Lysates from HEK293T cells expressing GFP-GJB2 were used as a source of fluorescently labeled GJB2. HEK293T ce...
Monolith NT.115:MST技术的代表仪器 Monolith NT.115是NanoTemper公司推出的一款采用MST技术的先进仪器。该仪器具有单次实验时间短(≤15分钟)、所需样品数量少(4-10μl)等优点,能够支持从离子、小分子到高分子量的蛋白复合体等多种类型的生物分子、化合物、纳米材料等分子间的相互作用研究。此外,NT.115还支持样品...
MST除了能精确检测生物分子间相互作用,通过计算解离常数(Kds),还能得到有关生物分子互作的其他参数,实现精确的定性分析。 NanoTemper Monolith NT.115 检测设备。下方为容纳 16 根毛细管的托盘。 三、案例介绍 (1)模拟体内蛋白-蛋白结合对比实验 实验目的:验证X 与 野生型YWT,X 与 突变型 YMu两组蛋白的结合能力差...
in 293T cells and then lysed the collected cells with IP lysis buffer. The affinity between the whole cell lysate and the MBI peptides (WT: APSEDIWKKFELLPTPPLSP, T58A: APSEDIWKKFELLPAPPLSP, T58D: APSEDIWKKFELLPDPPLSP) was detected by a Monolith NT.115 instrument (NanoTemper). The ...
Monolith NT.115主要参数: 可测定样品类型:离子、化合物、核酸、多肽、蛋白、糖类、脂质体、纳米颗粒、病毒等; 测定亲和力(KD值)范围:1 pM – mM; 测定样品的分子量范围:101- 107Da; 获得亲和力所需要的蛋白样品量小于0.5 μg; 一次可测定的样品数量:16个; ...
the Monolith His-Tag Labelling Kit RED-tris-NTA 2nd Generation (NanoTemper Technologies). Serially diluted bufalin compounds, with concentrations of 500 μM to 15 nM, were mixed with cell lysis solutions containing labelled E2F2, ZFP-GFP and GFP at room temperature and then loaded into Monolith ...
(WT: APSEDIWKKFELLPTPPLSP, T58A: APSEDIWKKFELLPAPPLSP, T58D: APSEDIWKKFELLPDPPLSP) was detected by a Monolith NT.115 instrument (NanoTemper). The concentration of MBI peptide was set as follows: 2.5 mM, 0.51 × 2.5 mM, 0.52 × 2.5 mM, 0.53 × 2.5 mM, 0.54 × 2.5 mM, 0.55 ...
The MST binding assay using NanoTemper monolith NT.115 was described previously (Qu et al., 2016, 2018). When purified protein was used as target, protein was labeled with Monolith Series Protein Labeling Kit (MO-L011, NanoTemper Technologies). When cellular protein was used for target protein...
The MST binding assay using NanoTemper monolith NT.115 was described previously (Qu et al., 2016, 2018). When purified protein was used as target, protein was labeled with Monolith Series Protein Labeling Kit (MO-L011, NanoTemper Technologies). When cellular protein was used for target protein...