NOTE: After the coupling of the first C-terminal amino acid of all peptides to the amino group of the membrane, the amino groups between the spots and likewise a number of the amino teams inside the spot areas do not form bonds with the amino acids. Make certain all the purification buffers are at 4 °C and carry out the protein purification within the chilly room to avoid protein denaturation. OBSERVE: This avoids the coupling of amino acids in the further cycles with the membrane instead of the rising peptide chain.
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