█ SDS-PAGE 5% 浓缩胶配方表:
组分名称 | 1ml | 2ml | 4ml | 4ml | 5ml | 6ml | 8ml | 10ml |
超纯水 | 0.68ml | 1.40ml | 2.10ml | 2.70ml | 3.40ml | 4.10ml | 5.50ml | 6.80ml |
30% Acrylamide 储液 | 0.17 ml | 0.33 ml | 0.50 ml | 0.67 ml | 0.83 ml | 1.00 ml | 1.30 ml | 1.70 ml |
1.0 M Tris-HCl (pH 6.8) | 0.13 ml | 0.25 ml | 0.38 ml | 0.50 ml | 0.63 ml | 0.75 ml | 1.00 ml | 1.25 ml |
10% SDS | 0.01 ml | 0.02 ml | 0.03 ml | 0.04 ml | 0.05 ml | 0.06 ml | 0.08 ml | 0.10 ml |
10% 过硫酸铵(APS) | 0.01 ml | 0.02 ml | 0.03 ml | 0.04 ml | 0.05 ml | 0.06 ml | 0.08 ml | 0.10 ml |
TEMED | 1.0 μl | 2.0 μl | 3.0 μl | 4.0 μl | 5.0 μl | 6.0 μl | 8.0 μl | 10.0 μl |
█ SDS-PAGE 分离胶配方表:
组分名称 | 5ml | 10ml | 15ml | 20ml | 25ml | 30ml | 40ml | 50ml | |
6% | 超纯水 | 2.6 ml | 5.3 ml | 7.9 ml | 10.6ml | 13.2ml | 15.9ml | 21.2ml | 26.5ml |
30% Acrylamide 储液 | 1.0 ml | 2.0 ml | 3.0ml | 4.0 ml | 5.0ml | 6.0ml | 8.0ml | 10.0ml | |
1.5 M Tris-HCl(pH 8.8) | 1.3 ml | 2.5 ml | 3.8ml | 5.0 ml | 6.3ml | 7.5ml | 10ml | 12.5ml | |
10% SDS | 0.05ml | 0.1 ml | 0.15ml | 0.2 ml | 0.25ml | 0.3 ml | 0.4 ml | 0.5 ml | |
10% 过硫酸铵(APS) | 0.05ml | 0.1 ml | 0.15ml | 0.2 ml | 0.25ml | 0.3ml | 0.4ml | 0.5 ml | |
TEMED | 4.0 μl | 8.0 μl | 12ul | 16.0μl | 20μl | 24μl | 32μl | 40μl | |
组分名称 | 5ml | 10ml | 15ml | 20ml | 25ml | 30ml | 40ml | 50ml | |
8% | 超纯水 | 2.3 ml | 4.6 ml | 6.9ml | 9.3 ml | 11.6ml | 13.9ml | 18.5 ml | 23.2ml |
30% Acrylamide 储液 | 1.3 ml | 2.7 ml | 4.0 ml | 5.3 ml | 6.7 ml | 8.0 ml | 10.7 ml | 13.3ml | |
1.5 M Tris-HCl(pH 8.8) | 1.3 ml | 2.5 ml | 3.8 ml | 5.0 ml | 6.3 ml | 7.5 ml | 10.0 ml | 12.5ml | |
10% SDS | 0.05 ml | 0.1 ml | 0.15 ml | 0.2 ml | 0.25 ml | 0.3 ml | 0.4 ml | 0.5 ml | |
10% 过硫酸铵(APS) | 0.05ml | 0.1 ml | 0.15 ml | 0.2 ml | 0.25 ml | 0.3 ml | 0.4 ml | 0.5 ml | |
TEMED | 3.0 μl | 6.0 μl | 9.0 μl | 12.0 μl | 15.0 μl | 18.0 μl | 24.0 μl | 30.0μl | |
组分名称 | 5ml | 10ml | 15ml | 20ml | 25ml | 30ml | 40ml | 50ml | |
10% | 超纯水 | 1.9 ml | 3.9 ml | 5.9ml | 7.9 ml | 9.9ml | 11.9ml | 15.8ml | 19.7ml |
30% Acrylamide 储液 | 1.7 ml | 3.3 ml | 5.0 ml | 6.7 ml | 8.3 ml | 10.0ml | 13.3ml | 16.7 ml | |
1.5 M Tris-HCl(pH 8.8) | 1.3 ml | 2.5 ml | 3.8 ml | 5.0 ml | 6.3 ml | 7.5 ml | 10.0ml | 12.5 ml | |
10% SDS | 0.05ml | 0.1 ml | 0.15 ml | 0.2 ml | 0.25ml | 0.3 ml | 0.4 ml | 0.5 ml | |
10% 过硫酸铵(APS) | 0.05ml | 0.1 ml | 0.15ml | 0.2 ml | 0.25ml | 0.3 ml | 0.4 ml | 0.5 ml | |
TEMED | 2.0 μl | 4.0 μl | 6.0 μl | 8.0 μl | 10.0 μl | 12.0 μl | 16.0 μl | 20.0 μl | |
组分名称 | 5ml | 10ml | 15ml | 20ml | 25ml | 30ml | 40ml | 50ml | |
12% | 超纯水 | 1.6 ml | 3.3 ml | 4.9 ml | 6.6 ml | 8.2 ml | 9.9 ml | 13.2 ml | 16.5 ml |
30% Acrylamide 储液 | 2.0 ml | 4.0 ml | 6.0 ml | 8.0 ml | 10.0 ml | 12.0ml | 16.0 ml | 20.0 ml | |
1.5 M Tris-HCl(pH 8.8) | 1.3 ml | 2.5 ml | 3.8 ml | 5.0 ml | 6.3 ml | 7.5 ml | 10.0 ml | 12.5 ml | |
10% SDS | 0.05ml | 0.1 ml | 0.15ml | 0.2 ml | 0.25 ml | 0.3 ml | 0.4 ml | 0.5 ml | |
10% 过硫酸铵(APS) | 0.05ml | 0.1 ml | 0.15ml | 0.2 ml | 0.25 ml | 0.3 ml | 0.4 ml | 0.5 ml | |
TEMED | 2.0 μl | 4.0 μl | 6.0 μl | 8.0 μl | 10.0 μl | 12.0 μl | 16.0 μl | 20.0 μl | |
组分名称 | 5ml | 10ml | 15ml | 20ml | 25ml | 30ml | 40ml | 50ml | |
15% | 超纯水 | 1.1ml | 2.3ml | 3.4ml | 4.6ml | 5.7ml | 6.9ml | 9.2ml | 11.5ml |
30% Acrylamide 储液 | 2.5 ml | 5.0ml | 7.5 ml | 10.0 ml | 12.5ml | 15.0ml | 20.0ml | 25.0ml | |
1.5 M Tris-HCl(pH 8.8) | 1.3ml | 2.5ml | 3.8ml | 5.0ml | 6.3ml | 7.5ml | 10.0ml | 12.5ml | |
10% SDS | 0.05 ml | 0.1ml | 0.15ml | 0.2ml | 0.25ml | 0.3 ml | 0.4 ml | 0.5ml | |
10% 过硫酸铵(APS) | 0.05 ml | 0.1ml | 0.15ml | 0.2ml | 0.25ml | 0.3 ml | 0.4 ml | 0.5ml | |
TEMED | 2.0 μl | 4.0 μl | 6.0 μl | 8.0 μl | 10.0μl | 12.0μl | 16.0μl | 20.0 μl |
▲注意
1. 超纯水:作为无杂质溶剂,提供均一的液相分散介质,保障凝胶聚合与电泳过程无外源干扰。
2. 30% Acrylamide 储液(丙烯酰胺 - 甲叉双丙烯酰胺):提供凝胶的单体与交联剂,聚合后形成三维多孔网状结构,为蛋白质分离提供分子筛效应。
3. 1.0 M Tris-HCl(pH 6.8)(仅浓缩胶用):构建浓缩胶的缓冲体系,维持 pH 稳态,与分离胶的 pH 梯度共同促成蛋白质的浓缩效应。
4. 1.5 M Tris-HCl(pH 8.8)(仅分离胶用):构建分离胶的高 pH 缓冲体系,为蛋白质电泳提供稳定的电荷环境,确保分离的准确性。
5. 10% SDS(十二烷基硫酸钠):阴离子去污剂,使蛋白质变性并结合负电荷,消除其天然电荷与构象差异,实现蛋白质按分子量的电荷密度均一化分离。
6. 10% 过硫酸铵(APS):自由基引发剂,在碱性条件下分解产生氧自由基,触发丙烯酰胺的自由基聚合反应。
7. TEMED(四甲基乙二胺):亲核性催化剂,加速 APS 的自由基生成,调控凝胶聚合速度
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