Purification of decoy proteins
The recombinant decoy proteins (TTc R1225E W1288A and LCHn) were
purified individually by NiSO4 IMAC as outlined above,
with the exception that 1 mL, gravity fed NTI-sepharose columns
(Thermofisher) were used. Additionally, 0.5 mM CHAPS was incorporated
into the sample buffer and an additional column wash incorporating 0.5
mM CHAPS was added. Wash and elution buffers included 20 mM and 300 mM
imidazole, respectively. Post-IMAC purification the samples were
concentrated, and buffer exchanged into PBS using Vivaspin20 columns (30
kDa or 50 kDa MWCO; Sartorious).