After cooling the cell culture on ice for 10 min, isopropyl β-d-1-thiogalactopyranoside (IPTG) was added to a final concentration of 0.1 mM and cells were incubated at 28 °C with shaking for additional 2.5~3 h. After harvesting cells by centrifugation, their lysis and protein...
Therefore, in this study we aimed to investigate a novel strategy that would allow the conversion of non-responsive polysaccharides into thermoresponsive polysaccharides with tuneable LCSTs. Through the functionalisation of dextran with alkylamide groups (isopropyl amide, diethyl amide, piperidinyl and ...
Then, isopropyl-thiogalactopyranoside (IPTG, Invitrogen) was added to a final concentration of 0.5 mM for 18 h to induce protein expression. Cells were harvested by centrifugation at 8500 rpm for 20 min at 4 °C. To prepare a total-cell lysate the pellet was frozen (−20 °C) for 24...
Expression of recombinant PRDX6 was induced with 1mM isopropyl b-d-thiogalactopyranoside (IPTG, Sigma). The culture was incubated for 20h at 18C, bac- teria were harvested by centrifugation (6000 g, 10 min, 4C) and resuspended in lysis...
Then, the expression of RsdA or RsdAΔC-terminal was induced with 0.5 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) at 16 °C for 20 h. The cells were harvested and lysed by freeze-thaw, and cell debris was removed by centrifugation. Recombinant RsdA or RsdAΔC-terminal ...
The crude product was dissolved in methylene chloride and precipitated with isopropyl alcohol. The wet product was dried under reduced pressure. Yield 6.8 g. NMR (d6-DMSO): 2.60 ppm (t, —CH2—COO—), 3.51 ppm (s, PEG backbone), 4.62 ppm (m, —CH2—O(C═O)—), 5.11 ppm (s, ...
(e.g. ethanol), isopropyl acetate or mixtures thereof, more preferably at between room and reflux temperature, in the presence of a suitable base (e.g. potassium carbonate) and an appropriate organic solvent (e.g. a lower alkyl alcohol, such as ethanol); or (b), for compounds of ...
coli, and induced with 0.2 mM of isopropyl β-D-1-thiogalactopyranoside (IPTG). The soluble DgAS proteins were then successfully purified with Ni2+ chelate affinity chromatography, as shown in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (Figure 1b). Figure 1. Production...
(N-ethyl-N-isopropyl) amiloride (EIPA) inhibited ATP-induced Na+fluxes by 95% and 72% at 40 μM, respectively [10]. Likewise, HMA also inhibited ATP-induced ethidium+uptake in human leukemic B cells by 50% at 20 μM, but concentrations > 100 μM were required for complete inhibition...
(OD600) of 0.6–0.8. One or more stable isotopes were supplemented to the growth media according to the NMR experimental requirements. Cells were cooled to 25 °C before the induction with 0.5 mM isopropyl β-D-thiogalatopyranoside (IPTG) at 25 °C overnight. Full deuteration of ...