Elsevier

Methods in Enzymology

Volume 72, 1981, Pages 296-303
Methods in Enzymology

[16] Protein determination in membrane and lipoprotein samples: Manual and automated procedures

https://doi.org/10.1016/S0076-6879(81)72018-4Get rights and content

Publisher Summary

This chapter describes the protein determination in membrane and lipoprotein samples. A variety of methods have proved to be effective in estimating the protein content of water-soluble samples. The procedure used is based on this modified Lowry method. It is found that by adding sodium dodecyl sulfate to the alkali reagent, samples can be assayed directly without prior solubilization or delipidation. An increase in the copper tartrate concentration facilitates quantitation of protein in the presence of sucrose and EDTA. Color formation depends mainly on reduction of the Folin-Ciocalteu reagent by protein-bound copper and proceeds in two distinct steps. The wavelength used routinely in the modified Lowry procedure was chosen as a compromise between increased absorption of the final blue reduction product with longer wavelengths and the practical limitations of most spectrophotometers. The absorption peak of the blue chromophore extends through much of the visible spectrum in a broad plateau and reaches maximum at 750 nm. The effectiveness and rapidity of this modified Lowry procedure as compared to the original Lowry procedure for assaying complex biological systems are also discussed.

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