Isoélectrofocalisation
L isoelectrofocalisation , or method of focusing electric, makes it possible to separate from the Protéines which differ only by only one load. One uses it in particular at the time of the two-dimensional electrophoresis, after having separated proteins according to their total electric charge in a gradient from pH in absence of Laurylsulfate of sodium (solvent conferring on proteins a negative charge). For the solubilization of proteins, one uses: a Detergent noncharged (Triton 100X), the Beta-mercaptoéthanol (breaking the bridges Disulfide) and the Urea (denaturing proteins without their conferring of load). One makes migrate proteins subjected to a gradient of pH in a tube of Gel of Polyacrylamide in their applying a Electric field. The proteins will present to the level of a certain pH, called not isoelectric a null total load (the protein is neutral). One thus separated proteins according to their isoelectric point.
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