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. 2002 Oct 28;99(23):15084–15088. doi: 10.1073/pnas.222561699

Fig 4.

Fig 4.

Chemical modification of proteins with nornicotine. SDS/PAGE and Western blot with mAb NIC6C12 of samples are shown. (a) BSA (lane 1), BSA + nornicotine and glucose (lane 2), HSA (lane 3), HSA + nornicotine and glucose (lane 4). (b) Chemoluminescent development of Western blot from human plasma; HSA + nornicotine and glucose (lane 1), nonsmoker sample A total protein concentrations of 2.5 mg/ml (lane 2), 5.0 mg/ml (lane 3), 10 mg/ml (lane 4); nonsmoker sample B total protein concentrations of 2.5 mg/ml (lane 5), 5.0 mg/ml (lane 6), 10 mg/ml (lane 7); smoker sample C total protein concentrations of 2.5 mg/ml (lane 8), 5.0 mg/ml (lane 9), 10 mg/ml (lane 10); smoker sample D total protein concentrations of 2.5 mg/ml (lane 11), 5.0 mg/ml (lane 12), 10 mg/ml (lane 13). Reactions were performed by incubating sterile solutions (0.2-μm filter) of glucose (200 mM), nornicotine (0.8 mM), and protein (0.8 mM) in phosphate buffer (200 mM, pH 7.4) in the dark at 37°C.