RFHR is an abbreviation of "radical-free and highly reducing" method, and an improved method of the 2D PAGE developed by Kaltschmidt and Wittmann.
On the contrary to pH gradient gel and concentration of proteins to the isoelectric point in O'Farrell's method, The RFHR 2D PAGE carries out definite pH electrophoresis at both the first and the second dimensions, and separates well proteins including basic proteins with difference of migration rate on highly reducing conditions.
These features of the RFHR2D PAGE brings about wonderful results to,and supplements the weak points of O'Farrell's method.
These features of the RFHR2D PAGE brings about wonderful results to,and supplements the weak points of O'Farrell's method.
Background of two dimensional of polyacrylamide gel electrophoresis
Analysis and separation of proteins in proteomics has been carried out by theO'Farrell's isoelectric point 2D PAGE.
However, O'Farrell's method has several serious weak points which includes low rate of gene identification from protein spots, low separation ability?for basic proteins and artificial splitting of isoelectric point during migration at the first dimension.Therefore, proteomics needs essential improvements of the method of applied 2D PAGE.
Analysis and separation of proteins in proteomics has been carried out by theO'Farrell's isoelectric point 2D PAGE.
However, O'Farrell's method has several serious weak points which includes low rate of gene identification from protein spots, low separation ability?for basic proteins and artificial splitting of isoelectric point during migration at the first dimension.Therefore, proteomics needs essential improvements of the method of applied 2D PAGE.
J.O'Farrell ( 1975, J. Biol. Chem., 250, 4007-4021 )
Akira Wada "Analysis of Escherichia coli ribosomal proteins by an improved two dimensional gel electrophoresis.
1. Detection of four new proteins" ( 1986, J. Biochem., 100, 1583-1594 )
Akira Wada "Analysis of Escherichia coli ribosomal proteins by an improved two dimensional gel electrophoresis.
2. Characterization of four new proteins" (1986, J. Biochem., 100, 1595-1605 )
Akira Wada "Analysis of Escherichia coli ribosomal proteins by an improved two dimensional gel electrophoresis.
1. Detection of four new proteins" ( 1986, J. Biochem., 100, 1583-1594 )
Akira Wada "Analysis of Escherichia coli ribosomal proteins by an improved two dimensional gel electrophoresis.
2. Characterization of four new proteins" (1986, J. Biochem., 100, 1595-1605 )
