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The remaining buffer was carefully removed and 3flag-tagged proteins were eluted using 90 l of flag elution buffer (20 mM HEPES-KOH (pH 7.9), 150 mM KCl, 1 mM DTT and 1 mg/ml 3flag peptide, Sigma-Aldrich, F4799)

In addition, copper can affect mitogen-activated protein kinase kinase 1 (MEK1) and mitogen-activated protein kinase kinase 2 (MEK2) and enhance their phosphorylation of ERK1 and ERK2 in a dose-dependent manner, further promoting tumor cell proliferation (85)

Markus Vogel, Head of Bioconjugation Research (Germany) "We needed a cyclic disulfide linker tailored to a specific antibody that would enhance conjugation efficiency without compromising immunoreactivity

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Then, the media was removed and cells were washed twice with PBS