对流免疫电泳缓冲液及电泳介质的筛选

Selection of Buffer Solution and Medium for Counter-immunoelectrophoresis

  • 摘要: 为找到对流免疫电泳中安全经济且效果好的电泳缓冲液以及电泳介质,从常用缓冲液中筛选出6种pH值在8.6的缓冲液进行电泳,分别用琼脂和琼脂糖作为电泳介质,观察电泳后抗原抗体形成白色沉淀线的情况。研究结果显示,6种缓冲液中Tris-HCl缓冲液效果最好,琼脂和琼脂糖凝胶均可产生明显的白色沉淀线,且随着抗原浓度的降低白色沉淀线逐渐变浅;巴比妥−巴比妥钠、TAE、Gly-NaOH和NaCl-NaOH缓冲液中琼脂糖作为电泳介质的效果均比琼脂好,TGS缓冲液效果最差。从而得出结论,Tris-HCl、巴比妥和巴比妥钠、TAE、Gly-NaOH和NaCl-NaOH缓冲液均可作为对流免疫电泳的缓冲液,但从安全与经济角度以及实验效果角度考虑Tris-HCl最好,TAE、Gly-NaOH次之。因此,免疫学实验课中将Tris-HCl作为对流免疫电泳的缓冲液。

     

    Abstract: In order to select a safe, economical and effective buffer solution and medium for counter-immunoelectrophoresis (CIEP), six buffer solutions with pH value of 8.6 were selected from common buffer solutions for electrophoresis, with agar and agarose used respectively as the electrophoresis media, to observe the formation of white precipitation line of antigens and antibodies after electrophoresis. The results showed that Tris-HCl buffer was the best of the six buffer solutions. Both agar and agarose gels produced significant white precipitation lines, and the white precipitators became shallower with the decrease of antigen concentration. The effects of agarose used as the electrophoresis media in barbiturate and barbiturate sodium, TAE, Gly-NaOH and NaCl-NaOH buffers were better than those of agar. The TGS buffer was the worst. It was concluded that Tris-HCl, Barbitone and barbitone sodium, TAE, Gly-NaOH and NaCl-NaOH buffer could all be used as buffer solution for counter immunoelectrophoresis. However, Tris-HCl was the best, followed by TAE and Gly-NaOH, in terms of safety, economy and experimental effect. Therefore, Tris-HCl will be used as buffer solution for counter-immunoelectrophoresis in the future immunological experiments.

     

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