不同的蛋白将具有不同的 Protein Thermal Shift 表达谱,各自具有独特的蛋白熔解曲线形状、斜率、信噪比和温度熔解范围(有关示例,请参见右图)。Protein Thermal Shift 软件通过两种方法从这些曲线生成一个或多个熔解温度值 (Tm):Boltzmann 派生的 Tm 和衍生曲线确定的 Tm,在不同检测条件下,用作曲线间的比较点并...
Applied Biosystems™ Protein Thermal Shift™ 试剂盒使用户可利用与暴露的疏水残基结合的染料,通过实时荧光定量 PCR 仪器来监测蛋白质的热稳定性,从而鉴定使目标蛋白稳定的缓冲液条件或筛选配体库以寻找与目标蛋白结合的化合物。节省样品筛查所花费的时间和费用 了解更多信息 ...
As a result, we developed the Protein ThermalShiftTM software v1.0 which utilizes theBoltzmann equation to calculate the Tm of theprotein from the fluorescence melt plot.Comparisons can then be made between Tmvalues obtained using a range of bufferconditions, addition of different ligands, or...
5. Correlate changes in protein stability or ligand binding to changes in Tm. The Protein Thermal Shift Software calculates the Tm from each fluorescence profile using the Boltzmann method (from a plot of fluorescence intensity vs. temperature) and the derivative method (from a plot of d(...
study to identify buffer conditions that increase thermal stability of the protein, then repeat the original study using the new buffer conditions. i am getting an error message when i try to open my *.eds data ...
The melting temperatures (Tm) were taken from the minimum derivative plot (c), plotted as the negative of the first derivative of the normal fluorescent curve, converted by Protein Thermal Shift software. The Tm values were determined as the mean + s.d. of n = 3 replicates. d, e,...
respectively for 5-25 min. The results of live cells (using excitation wavelength of 490 nm) and dead cells (using excitation wavelength of 528 nm) were recorded by in C2 + confocal laser scanning microscope (Nikon, Japan) with NIS-Elements AR 4.20 software. For the cell apoptos...
The fluorescence intensity was measured during a temperature gradient from 25 to 95 °C at a constant rate of 0.05 °C/s, and protein melting temperatures were calculated based on a Boltzmann function fitting to experimental data, as implemented in the Protein Thermal Shift Software (Applied ...
(8X), and 5 μl of Protein thermal shift buffer was dispensed in each well. The PCR plates were then sealed with optical seal and centrifuged to remove the air bubbles in the mixture. Thermal scanning (25–99.9 °C at 0.05 °C/s) was done using real-time PCR system (...
(Fig.2a, this value is smaller than that in our previous report since we used the protein with a higher purity this time). The cellular thermal shift assay (CETSA) was carried out to test the interaction of LRPPRC and GAA in cells32, which showed that GAA could increase the thermal ...