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The SHT not only delivers performance enhancement for event querying in the trace [15], but allows to dissociate the analysis step from the trace itself.
This procedure allows to dissociate Hydra tissue into a suspension of viable single cells.
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MiR-608 or -132 oligos were diluted in buffer (serial two-fold dilutions, 0.3125, 0.625, 1.25, 2.5, 5 and 10 μ m) and injected over the flow cells for 2 min at 10 μl/min, with 5-min association and 5-min dissociation, except for the highest concentration that was allowed to dissociate for 1 h.
After the end of each injection, antibodies were allowed to dissociate for 20min (1200sec) and then the chip was regenerated with 100mM HCl (BIAcore).
PriL-CTD protein in buffer F containing 0.005% P20 was injected at several concentrations for 60 seconds and allowed to dissociate for 60 seconds.
Complexes were allowed to dissociate for approximately three minutes, after which a solution of wtDR1/HAY308A complexes (∼2.5 µM) pre-incubated with SEH or SEA at a 1∶1 molar stoichiometry was injected over the surface of flow cell 1 or 2 respectively for another ten minutes.
The complex was allowed to dissociate for 800 s.
S20 was allowed to dissociate by incubation of the sensors in PBS with 0.1% Tween-20 and 10% DMSO.
Increasing concentrations of CFHR312 or fH67 were injected over the flow channels at 60 µl/min for 240 s and allowed to dissociate for 1200 s.
For simplicity, in our model, WIPI1/2 and PtdIns3P are not allowed to dissociate from a ternary complex of ATG9, WIPI1/2 and PtdIns3P.
We observed K GDP values for EF-Tu that ranged from 30 to 75 nM in the absence of EF-Ts at the temperatures in which the EF-Tu·GDP complex was allowed to dissociate (Table 2).
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