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This lack of activity may be due to loss of hydrophobic interactions, but it is also possible that the methyl group affects the conformation of the amide and leads to disruption of the H-bond between the carbonyl and T336

For protein quantification, the remaining 50 L of the cell suspension underwent 3 freeze-thaw cycles using liquid nitrogen, followed by centrifugation at 13,800 g for 10 min

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Binding protein interactions: Standard IGF-1 binds IGFBPs (insulin-like growth factor binding proteins) strongly limiting free active peptide

The centrifugation process was performed after 10 min at 400g for 10 min, followed by separation and disposal of the plasma and buffy coat

Marklund SL, Holme E, et al