EVALUATION OF ISOELECTRIC-FOCUSING RUNNING CONDITIONS DURING TWO-DIMENSIONAL ISOELECTRIC-FOCUSING SODIUM DODECYL SULFATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS - VARIATION OF GEL PATTERNS WITH CHANGING CONDITIOS AND OPTIMIZED ISOELECTRIC-FOCUSING CONDITIONS
EVALUATION OF ISOELECTRIC-FOCUSING RUNNING CONDITIONS DURING TWO-DIMENSIONAL ISOELECTRIC-FOCUSING SODIUM DODECYL SULFATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS - VARIATION OF GEL PATTERNS WITH CHANGING CONDITIOS AND OPTIMIZED ISOELECTRIC-FOCUSING CONDITIONS
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DOI：
10.1016/0003-2697(84)90783-8
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发表时间：
1984-01-01
影响因子：
2.9
通讯作者：
HERSHEY, JWB
中科院分区：
文献类型：
--
作者：
DUNCAN, R;HERSHEY, JWB
系统地改变5个主要等电聚焦（IEF）参数：伏特小时、丙烯酰胺、NaOH和H3PO4浓度以及平衡时间，以确定各参数对[人宫颈癌HeLa细胞蛋白]二维IEF/十二烷基硫酸钠-聚丙烯酰胺凝胶电泳模式的影响，并优化IEF条件。每个参数的改变都会影响凝胶模式，经常导致在条件之间斑点识别的不确定性。结果强调了内部分析一致性的必要性，并表明如果采用不同的IEF条件，实验室之间的凝胶模式比较可能会很复杂。系统评价表明，当NaOH和H3PO4浓度增加（分别为50和25 mM），运行时间为10,000 V-h或更长时，可以获得最佳的图案。
Five major isoelectric focusing (IEF) parameters, volt-hours, concentrations of acrylamide, NaOH, and H3PO4 and equilibrium time, were systematically varied to determine the effect of each on 2-dimensional IEF/sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel patterns [of human cervical carcinoma HeLa cell proteins] and to optimize IEF conditions. Alterations in each parameter affected the gel pattern, frequently causing uncertainty in the identification of spots between conditions. The results emphasize the need for internal analytical consistency, and indicate that gel pattern comparisons between laboratories can be complicated if different IEF conditions are employed. The systematic evaluation indicated that optimized patterns were obtained when increased concentrations of NaOH and H3PO4 (to 50 and 25 mM, respectively) and run durations of 10,000 V-h or longer were used.