4.5. Purification of Recombinant Irisin from IBs

The Ni–NTA column was equilibrated with equilibration buffer, and the collected supernatant was loaded on the equilibrated Ni–NTA column. Then, the column was washed with washing buffer (equilibration buffer + 20 mM imidazole) and the desired protein was eluted with elution buffer (buffer Y) with different concentrations of imidazole. The fractions of the eluted irisin protein were collected and checked for homogeneity on SDS-PAGE.