HTLV-1 CA, CA-NTD, and CA-CTD were separately concentrated in Pierce concentrator tubes (10 MWCO for CA and CA-NTD, and 3 MWCO for CA-CTD). Monomeric HTLV-1 CA, CA-NTD, and CA-CTD were diluted to 1 mg/mL concentration in both dialysis buffer and under assembly conditions (50 mM MES, 200 mM NaCl, pH6) in a volume of 10 μl and placed in Prometheus Series High Sensitivity Capillaries (NanoTemper, Cat. no. PR-C006).
For measurement of assemblies the proteins were first placed in assembly buffer (50 mM MES, 200 mM NaCl, pH6) with a final protein concentration of 16 mg/mL and were incubated at 4°C for 4h and 26 °C overnight. Again, a volume of 10 μl was then placed in Prometheus Series High Sensitivity Capillaries (NanoTemper, Cat. no. PR-C006).
For measurement of assemblies the proteins were first placed in assembly buffer (50 mM MES, 200 mM NaCl, pH6) with a final protein concentration of 16 mg/mL and were incubated at 4°C for 4h and 26 °C overnight. Again, a volume of 10 μl was then placed in Prometheus Series High Sensitivity Capillaries (NanoTemper, Cat. no. PR-C006).