WebProtein thermostability has been investigated by two approaches. (A) Computational. To study the relationship between thermostability and conformational characteristics of … WebOf particular interest is the fluorescence-detector size-exclusion chromatography-based thermostability assay (FSEC-TS). By simple heating and FSEC, the generally applicable method allows rapid assessment of the thermostability of GFP-fused membrane proteins without purification. Here we describe the experimental details and some typical ...
Fluorescence-detection size-exclusion chromatography utilizing nan…
WebSep 7, 2024 · For FSEC-based thermostability assays, 50 μL of cell lysate was heated in a PCR machine for 20 min, followed by 4°C incubation for 10 min. The heated samples were clarified by centrifugation before being loaded onto a ZenixC-300 column in a Shimadzu or Agilent HPLC machine equipped with a fluorescence detector for signal monitoring at the … WebDec 10, 2024 · Next, we tested the robustness of TGP for FSEC-TS assay using the ultra-stable PlsY from Aerocus aquifex ( aa PlsY, 20.9 kDa, 7 transmembrane helices), which has a half-life of 30 min at 90 °C rotary clubs in portugal
Fluorescence-Detection Size-Exclusion Chromatography-Based ...
Web6 3.0 SCOPE OF THE FSA Figure 1 below provides an overview of this safety lifecycle model: IEC61511 SIS S afety Lifecycl e P has es an d Funct ional Saf ety WebApr 20, 2024 · (ii) Thermostability assay. The thermostability of a detergent-solubilised GPCR depends upon the type of detergent used and whether the receptor is either ligand-free, agonist-bound or antagonist-bound [35, 36]. In addition, the receptor stability tends to be increased by an increase in affinity and/or decrease in the off-rate of the ligand ... WebDec 10, 2024 · The stability assay for hSI by tryptophan-FSEC was carried out exactly the same way as for ttSI except that the solubilization and purification steps were performed … stoughton extended day address