Total proteins in breast cancer rat tissues were isolated with
RIPA buffer (Thermo, 89900) containing 25 mM Tris–HCl (pH 7.6),
150 mM NaCl, 1% NP-40, 1% sodium deoxycholate and 0.1% SDS.
The protease inhibitor was added in to protein solution and protein
was ready for use or kept at 40 °C.