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Interestingly, the residues that had significant effects on GLP-1R internalisation while mutated to alanine were all non-polar branched amino acids, with some of these also predicted to cause changes in the cavity in which they reside

This involved adding a lipid-ethanol solution to a plasmid solution in 25 mM citrate buffer (pH 4.0), followed by buffer replacement with PBS through diafiltration using Amicon Ultra Centrifuga Filter (10 kDa MWCO, Millipore)

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