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Interference of salts used on aqueous two-phase systems on the quantification of total proteins

Author:
Simone Maria Golunski, Luisa Sala, Marceli Fernandes Silva, Rogério Marcos Dallago, Jéssica Mulinari, Altemir José Mossi, Adriano Brandelli, Susana Juliano Kalil, Marco Di Luccio, Helen Treichel
Source:
International journal of biological macromolecules 2016 v.83 pp. 30-33
ISSN:
0141-8130
Subject:
absorbance, enzyme activity, inulinase, metals, potassium phosphates, proteins, sodium chloride, sodium citrate, sodium nitrate
Abstract:
In this study the interference of potassium phosphate, sodium citrate, sodium chloride and sodium nitrate salts on protein quantification by Bradford's method was assessed. Potassium phosphate and sodium citrate salts are commonly used in aqueous two-phase systems for enzyme purification. Results showed that the presence of potassium phosphate and sodium citrate salts increase the absorbance of the samples, when compared with the samples without any salt. The increase in absorptivity of the solution induces errors on protein quantification, which are propagated to the calculations of specific enzyme activity and consequently on purification factor. The presence of sodium chloride and sodium nitrate practically did not affect the absorbance of inulinase, probably the metals present in the enzyme extract did not interact with the added salts.
Agid:
5293768