Enzymatic Metallization of Protein A/G

For the initial
investigation of the metal deposition, recombinant protein A/G (ThermoFisher
Scientific) was immobilized as the antigen. This was incubated for
60 min with 200 nM mouse anti-human IgG Fc-Horse Radish Peroxidase
(HRP) (SouthernBiotech) in 100 μL volume. Beads were washed
3 times in deionized water before the silver deposition was performed
with EnzMet (Nanoprobes). 50 μL each of EnzMet solutions A,
B, and C were incubated with beads for 4, 4, and 8 min, respectively.
Note that these times are obtained from assay optimization performed
in our prior work with enzymatic metallization.^{9 (link)}