血清蛋白醋酸纤维膜电泳缓冲液的筛选

Screening of Serum Protein Cellulose Acetate Membrane Electrophoresis Buffers

旨在筛选适合血清蛋白醋酸纤维膜电泳的最佳电泳缓冲液。方法:选择6种常见电泳缓冲液,采用醋酸纤维膜为支持物分离家兔血清蛋白,确定其电泳最佳电压;在相同实验条件下,观察使用频次和放置一定时间对各种缓冲液稳定性的影响,并对比图谱分离蛋白质区带的数量及清晰度,定量分析蛋白质区带膜条扩展的总距离、点样区距第一条带远端、最远区带与最近区带的距离。结果表明在电压条件为70 V时,各种缓冲液电泳分离血清蛋白均能达到较好水平,其中出现5条清晰区带的概率,Tris-磷酸盐缓冲液为66.66%,巴比妥缓冲液为65.22%、Tris-盐酸缓冲液为8.33%;各缓冲液在使用5次以内、室温条件放置6个月后均对缓冲液稳定性影响不大;图谱区带数量和清晰度、区带分散程度定量分析表明Tris-磷酸、巴比妥缓冲液效果最好,Tris-盐酸缓冲液次之,磷酸盐、硼酸盐缓冲液较差。结论:6种缓冲溶液中,以图谱分离5条蛋白质区带为准,Tris-磷酸盐缓冲液分离血清蛋白图谱区带清晰,与巴比妥缓冲液效果相当。

The aim is to screen the best electrophoresis buffer suitable for serum protein cellulose acetate membrane electrophoresis.Methods:Six common electrophoresis buffers were selected and cellulose acetate membranes were used as supports for the separation of rabbit serum proteins to determine their optimal voltage for electrophoresis;under the same experimental conditions,the effects of frequency of use and placement for a certain period of time on the stability of the various buffers were observed,and the number and clarity of the protein bands separated by comparison plots were quantified by analyzing the total distance of the membrane strip extension of the protein bands,the distance of the spot sample area from the distal end of the first band,and the distance between the farthest band and the nearest band.The results showed that at a voltage of 70 V,the electrophoretic separation of serum proteins in all buffers was good,and the probability of five clear bands was 66.66%for Tris-phosphate buffer,65.22%for barbiturate buffer and 8.33%for Tris-hydrochloric acid buffer;the stability of the buffers was not significantly affected after 6 months at room temperature within 5 times of use.The quantitative analysis of the number and clarity of the bands and the dispersion of the bands showed that Tris-phosphate and barbiturate buffers were the most effective,followed by Tris-hydrochloric acid buffer,and phosphate and borate buffers were less effective.Conclusion:Of the six buffer solutions,the Tris-phosphate buffer separated serum proteins with clear bands,based on the graphical separation of five protein bands,which was comparable to that of the barbiturate buffer.