Method for improving measurement accuracy through two-dimensional protein electrophoresis

Abstract

The invention provides a method for improving experiment accuracy by an isoelectric focusing method of two-dimensional protein electrophoresis, in particular a method for calculating electric energy needed by salt and protein focusing by measuring electric conductivity of a protein object to be measured. The method is characterized in that: electric energy needed by the protein focusing and salt mobility is calculated through a group of formulas in environments of different electric conductivities of the salt, the protein weight, the pH gradient gel length and the range of the pH value respectively, proper electric energy of the experiment of the individual protein object to be measured can be provided in the isoelectric focusing method so as to present an optimal protein focusing result,and the influence of colloidally protein focusing due to insufficient or over-high electric energy is avoided.

Description

technical field [0001] The present invention provides a method for improving the measurement accuracy of two-dimensional protein electrophoresis, especially a method for evaluating the salt content by measuring the electrical conductivity of the protein to be tested, and deriving the salt swimming and protein focusing required by the calculation formula electrical energy. Background technique [0002] Since the 1980s, bioengineering has become an important symbol of science and technology. Many important studies have relied on electrophoretic separation technology, which has become an indispensable experimental method in chemistry, molecular biology and related industries; The charge depends on the pH of the environment. Electrophoretic separation technology needs a medium as its place. Nowadays, semi-solid colloids are mostly used, mainly polyacrylamide gel as the electrophoretic support. On this basis, SDS-polyacrylamide is developed. Gel electrophoresis, isoelectric focu...

AI Technical Summary

Problems solved by technology

[0004] In the biological sample of the experiment, in addition to protein molecules, the colloid also includes a variety of salts. Because the salts also have the characteristics of electrolytes, a part of the voltage and electric energy is consumed in the electrophoresis and will be used for electrolytic salt swimming. , because the salt content in the sample is related to the experiment and biological characteristics, it cannot be accurately estimated; therefore, when operating the isoelectric focusing method of the two-dimensional electrophoresis analysis method, it is generally impossible to evaluate the electrical energy consumed by the salt, and it is also impossible to accurately measure the calculated sample. In isoelectric focusing electrophoresis, if the electric energy of the given protein molecules is insufficient, the isoelectric focusing method cannot produce good protein focusing on the isoelectric focusing colloid of the test piece If the electric energy provided is too large, the protein molecules on the test piece will swim away from the colloid and form over-focusing; therefore, most laboratories use protein Precipitation and extensive washing to remove salts in the protein sample, but at the same time many important proteins may be lost; even some specific protein samples cannot be effectively precipitated in the process

Images