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After three washes, 5 min each, in TBST (TBS with 0.1% Tween 20), the secondary antibody (goat anti-mouse-HRP, Cell Signaling Technology 1:2000 dilution) was incubated in TBST with 5% non-fat dry milk for 1 h at room temperature followed by four 5 min washes in TBST

R., Kinter, M

Because cardiovascular issues can be life-threatening, any sudden onset of these symptoms, especially with chest pain or pressure, should be treated as an emergency