We have developed a method for intact mass analysis of detergent- solubilized and purified integral membrane proteins using liquid chromatography-mass spectrometry (LC-MS) with methanol as the organic mobile phase. Membrane proteins and detergents are separated chromatographically during the isocratic stage of the gradient profile from a 150-mm C3 reversed-phase column. The mass accuracy is comparable to standard methods employed for soluble proteins; the sensitivity is 10-fold lower, requiring 0.2-5 μg of protein. The method is also compatible with our standard LC-MS method used for intact mass analysis of soluble proteins and may therefore be applied on a multiuser instrument or in a high-throughput environment. © 2010 Elsevier Inc. All rights reserved.
CITATION STYLE
Berridge, G., Chalk, R., D’Avanzo, N., Dong, L., Doyle, D., Kim, J. I., … Gileadi, O. (2011). High-performance liquid chromatography separation and intact mass analysis of detergent-solubilized integral membrane proteins. Analytical Biochemistry, 410(2), 272–280. https://doi.org/10.1016/j.ab.2010.11.008
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