At the end of the DNA injection, unreacted carboxyl groups were inactivated by injection of selection buffer twice for a total of twenty minutes, followed by a regeneration cycle with 45 mM glycine and 100 mM sodium hydroxide in 5% ethanol (regeneration buffer) for 30 seconds [ 36, 37].
The heat change induced by the injection of buffer into the lysozyme solution and that induced by the injection of TbTIM into buffer were subtracted from the experimental (TbTIM into lysozyme); the data are in Fig. 5B.
The resident peritoneal macrophages were obtained by injection of cold phosphate buffer solution (PBS) 1x (5 mL, 3x) into euthanized mice abdominal cavity.
Protein binding was evaluated during an association phase (0 300 sec), which was followed by a dissociation phase (injection of buffer only, 300 3700 sec).
Bound peptide was removed by injection of 5 μl regeneration buffer (25 mM NaOH, 0.002% SDS) at 10 μl/min.
The rapid preservation of the effects of altered gravity on the gene expression in the U937 cells was achieved by injection of 50 mL of RLT buffer (Qiagen, Germany), a lysis buffer immediately lysing cells and tissues prior to RNA isolation.
After measuring the baseline with HBS-EP buffer (pH 7.4), association with Protein A was analyzed by injection of Fc samples (HBS-EP buffer), followed by analysis of the dissociation of the Fc proteins from Protein A by injection of HBS-EP buffer only.
They were further corrected by subtraction of the curve that was obtained after injection of buffer alone.
For all experiments, a control flow cell was prepared by injection of 10 mM sodium acetate buffer, pH 4.1, directly after activation, followed by blocking with 1 M ethanolamine, pH 8.5.
The process is quite simple and only needs three steps: buffer injection only, sample and buffer at the same flow rate, and a second injection of buffer only.
