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When 1 μL of a 10 μmol L−1 solution in buffer is injected at a flow rate of 1 μL min−1 (165 s exposure time), the injected protein is completely digested, as was observed with UV detection (data not shown).
Subsequently, buffer is injected for the dissociation step.
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After allowing the flowcell surfaces to equilibrate with HBS-EP running buffer, solutions of varying concentrations of TGFβ1, diluted in HBS-EP buffer, were injected using kinetic analysis injection protocols through Biacontrol software.
Subsequently, 20 mL of collagenase (100 μg/mL dissolved in D-hanks buffer) were injected.
The mix of the crude enzyme and loading buffer was injected into the gel.
Proteins in the reaction buffer were injected into the chamber to measure binding to DNA substrate.
For leakage measurements, a small volume of protein solution in Tris buffer was injected into a quartz cuvette containing the liposome suspension under constant stirring to obtain the lipid/protein ratio (L/P) = 100/1.
Accordingly, we examined the heat changes that occur when 15 µl of buffer was injected into cells that contained different lysozyme concentrations (inset Fig. 5); the aim was to find a condition in which the signal induced by TbTIM was distinguishable from that induced by buffer alone.
Cold 2.5 % glutaraldehyde in phosphate buffer was injected into a lateral ventricle for 2 min.
PBS buffer was injected into the tube at a speed of approximately 0.01 MPa/s (75 mm Hg/s) until failure.
As a reference, HBS-EP running buffer was injected and followed by the secondary scFv as above.
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