A simple protocol adaptation to influence the tryptic digest could thus be to change the buffer conditions, e.g. to a buffer as used in eFASP (Supplementary protocol 4).
With the same 10 mM Tris buffer as used in Figure 3C,D, the probe's spectral progression and pH change are much slower in this chamber than in the other one mentioned above, reflecting a difference in the oxygen exchange rate that is allowed by the corresponding port dimensions.
The appearance of microcracks in cerium oxide (CeO2) buffer layers, as used in buffer layer architectures for coated superconductors, indicates the presence of stress between this buffer layer and the substrate.
All experiments were performed in exactly the same buffer compositions as used in the manuscript: 1) NMR dilution titrations provided experimental evidence for a very weak CLR01 dimer formation (Ka = ∼60M-1) in PBS buffer (10 mM sodium phosphate, 137 mM NaCl, 2.7 mM KCl, pH 7.4), with ∼5% CLR01 dimers present at 500 µM.
One part of cells (wet mass) was suspended in one volume of buffer (the same as used in enzyme assay) at usual concentrations from 10 to 50 mM containing 0.2 mg DNase I per mL of cell suspension.
Consequently, we opted to use 1 M GA loading buffer and 1% ammonia elution buffer (as also used in a manual method) for all TiO2 and Ti-IMAC automated phosphopeptide enrichments.
The next morning, slides were washed in PBS (3x, 5 min each), and incubated with a Cy3-conjugated affinity purified donkey anti-rabbit secondary antibody diluted 1∶500 in the same buffer as used for dilution primary antibody, for 2 hours at room temperature.
Different food matrices as well as buffers were used in these reports: beef, milk, egg [ 22], infant formula [ 14, 24], orange juice, apple juice [ 21], pancake mix, peanut butter [ 24], phosphate buffered saline (PBS) [ 22, 24], sodium phosphate/sodium acetate buffer [ 13, 25], and KCl buffer [ 23].
We chose Ni2+ Sepharose 6 Fast Flow and Q Sepharose Fast Flow beads according to the characteristics of the pET-22b plasmid and the CC10 protein as well as the elution buffer used in affinity chromatography purification.
The surface charge of the particles was examined via ζ-potential measurements (Zetasizer – Malvern) in a 0.001 M KCl electrolyte with a pH adjusted to 6.8 with 0.01 M NaOH and 0.01 M HCl (same pH as in the buffer solution used in the drug release measurements described below).
Even though pH 5 sodium acetate buffer was used in all reactions as suggested by Bailey et al.[ 14], this buffer was not able to establish an ideal pH for the reaction to occur, as the lignin is extremely basic in nature.
