Forced protein oxidation was introduced to the samples (30 μg, 1 mg/mL) by diluting with an equal volume of 0.1% H2O2 (Merck, 107,209) and incubation at room temperature. After 0, 6, and 24 h a 10 µL aliquot was taken and the oxidation reaction was stopped by buffer exchange to 25 mM NH4HCO3 (Merck, 101131), pH 7 with Amicon filter devices (Merck, UFC503096), respectively. To force protein deamidation 30 µg sample was buffer exchanged to 25 mM NH4HCO3 (Merck, 101131), pH 10 using Amicon filter devices (Merck, UFC503096). Subsequently, the sample volume was adjusted to 30 µL and incubated at 37°C. To stop the deamidation reaction the sample was buffer exchanged to NH4HCO3, pH 7 as described previously in the oxidation workflow.
Amicon filter devices
Manufactured by Merck Group
Amicon filter devices are laboratory equipment used for separation and purification of macromolecules, such as proteins and nucleic acids. These devices utilize membrane-based filtration to selectively retain or pass specific components of a sample based on their molecular weight. The core function of Amicon filter devices is to facilitate the concentration, desalting, or buffer exchange of macromolecular solutions.
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Lab products found in correlation
2 protocols using amicon filter devices
1
Oxidation and Deamidation of Proteins
2
Oxidation and Deamidation of Proteins
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Forced protein oxidation was introduced to the samples (30 μg, 1 mg/mL) by diluting with an equal volume of 0.1% H2O2 (Merck, 107,209) and incubation at room temperature. After 0, 6, and 24 h a 10 µL aliquot was taken and the oxidation reaction was stopped by buffer exchange to 25 mM NH4HCO3 (Merck, 101131), pH 7 with Amicon filter devices (Merck, UFC503096), respectively. To force protein deamidation 30 µg sample was buffer exchanged to 25 mM NH4HCO3 (Merck, 101131), pH 10 using Amicon filter devices (Merck, UFC503096). Subsequently, the sample volume was adjusted to 30 µL and incubated at 37°C. To stop the deamidation reaction the sample was buffer exchanged to NH4HCO3, pH 7 as described previously in the oxidation workflow.
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