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doi.org/10.1530/eje.0.1390552 Kojima, M., & Kangawa, K

The cells were fixed in 4% PFA in PBS for 30 min, permeabilized with 0.2% Triton X-100 in PBS for 5 min, treated with 2 M HCl for 20 min and neutralized with 0.1 M sodium tetraborate for 5 min