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(b) 1 was synthesized using standard fluorenyl-methoxy carbonyl protocols on PAP Tenta-Gel (Rapp Polymere). Cleavage by TFA and deprotection yielded a linear PEG 3000 covalently bound to the carboxyl terminus of the peptide. Peptide purity, determined by synthetic coupling yields and reverse phase HPLC of CNBr cleavage products, was >96%. Molecular masses of all peptides were verified by MALDI-TOF mass spectroscopy.
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3 (pH ≈ 3.0). The samples were mixed, centrifuged for 15 min at 14,000g and incubated for 1 h. The mixtures were then diluted to 1 mM in phosphate buffer at pH 5.7 and allowed to stand at room temp for 3 days. Samples were applied to a glow discharge 400 mesh carbon-coated support film, followed by staining with 1% uranyl acetate. Micrographs were recorded using Philips EM 300 at magnifications of 100,000, 45,000, and 10,000.
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A is Avogadro's number.
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