Troitsky G V, Zav'yalov V P, Kirjukhin I F, Abramov V M, Agitsky G J
Biochim Biophys Acta. 1975 Jul 21;400(1):24-31. doi: 10.1016/0005-2795(75)90123-3.
A new method of producing a stable pH gradient in buffer solutions is suggested, obtained by the concentration gradient of a nonelectrolyte in buffer solutions as a result of the gradual change in the dielectric properties of the solution. The maximal concentrations of nonelectrolyte which do not influence the protein configuration allow a pH gradient with a range of two pH units to be produced. It is suggested that the properties of some polyols (i.e. glycerol or mannitol) be used to change the pH of the borate buffer for the production of greater pH gradient with a range of up to 4-5 pH units. Creating the gradient of concentration of polyols, one can obtain a pH gradient in borate buffer solutions. Though the polylydroxyl compound-borate complexes posses mobility in an electric field, a stable pH gradient can be achieved during 12 days of electrophoresis. The isoelectric focusing of haemoglobin, human serum albumin and immunoglobulins was carried out in both systems suggested. These findings were compared with isoelectric focusing in Ampholines. There was a good agreement between the methods compared. The possible differences are discussed.
提出了一种在缓冲溶液中产生稳定pH梯度的新方法，该方法是通过缓冲溶液中非电解质的浓度梯度实现的，这是由于溶液介电性质的逐渐变化导致的。不影响蛋白质构象的非电解质最大浓度可产生范围为两个pH单位的pH梯度。建议利用某些多元醇（如甘油或甘露醇）的性质来改变硼酸盐缓冲液的pH值，以产生范围高达4 - 5个pH单位的更大pH梯度。通过创建多元醇浓度梯度，可以在硼酸盐缓冲溶液中获得pH梯度。尽管多羟基化合物 - 硼酸盐络合物在电场中具有迁移率，但在12天的电泳过程中仍可实现稳定的pH梯度。在上述两种系统中均对血红蛋白、人血清白蛋白和免疫球蛋白进行了等电聚焦。将这些结果与在两性电解质中的等电聚焦进行了比较。所比较的方法之间具有良好的一致性。还讨论了可能存在的差异。
