SDS-PAGE凝胶配制试剂盒
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产品编号 P0012A
数量
¥ 163.00
产品包装:
可制30-50块胶
产品简介
使用说明
说明书下载
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碧云天生产的SDS-PAGE凝胶配制试剂盒提供了配制SDS-PAGE凝胶所需的各种试剂,用户只需自备制胶器具和蒸馏水,即可配制PAGE胶(即聚丙烯酰胺凝胶)。
SDS-PAGE凝胶配制试剂盒不仅可用于配制SDS-PAGE凝胶,也可用于配制非变性(native)PAGE凝胶。
本试剂盒约可配制30-50块常规大小的PAGE胶。
包装清单:

产品编号 产品名称 包装
P0012A-1 30% Acr-Bis (29:1) 100ml
P0012A-2 1M Tris-HCl, pH8.8 100ml
P0012A-3 10% SDS 5ml
P0012A-4 凝胶聚合催化剂 0.5g
P0012A-5 TEMED 0.5ml
P0012A-6 1M Tris-HCl, pH6.8 15ml
说明书 1份

保存条件:
1M Tris-HCl,pH8.8、10% SDS、凝胶聚合催化剂和1M Tris-HCl,pH6.8室温保存。30% Acr-Bis(29:1)和TEMED 4℃避光保存。凝胶聚合催化剂加水配制成10%溶液后,分装成小管-20℃保存,通常半年内有效。
注意事项:
凝胶聚合催化剂配制成10%溶液后,应当-20℃保存。同时应尽量减少室温存放时间,以防失效。
TEMED易挥发,使用后请盖紧瓶盖。另外凝胶凝聚的速度和温度及光照关系密切,可通过适当调节凝胶聚合催化剂和TEMED的用量,控制在不同的室内环境下凝胶凝聚的速度。
TEMED易燃,有腐蚀性,操作时请小心,并注意有效防护以避免直接接触人体或腐蚀其他物品。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
1.10%凝胶聚合催化剂的配制:例如称取0.1g凝胶聚合催化剂,用水溶解,并定容至1ml,即为10%凝胶聚合催化剂。
2.根据目的蛋白的分子量大小选择合适的凝胶浓度,再按照下面的表格配制SDS-PAGE的分离胶(即下层胶):
不同浓度的SDS-PAGE分离胶的最佳分离范围:

SDS-PAGE分离胶浓度 最佳分离范围
6%胶 50-150kD
8%胶 30-90kD
10%胶 20-80kD
12%胶 12-60kD
15%胶 10-40kD

成分 配制不同体积SDS-PAGE分离胶所需各成分的体积(毫升)
6%胶 5 10 15 20 30 50
蒸馏水 2.0 4.0 6.0 8.0 12.0 20.0
30%Acr-Bis(29:1) 1.0 2.0 3.0 4.0 6.0 10.0
1M Tris, pH8.8 1.9 3.8 5.7 7.6 11.4 19.0
10%SDS 0.05 0.1 0.15 0.2 0.3 0.5
10%凝胶聚合催化剂 0.05 0.1 0.15 0.2 0.3 0.5
TEMED 0.004 0.008 0.012 0.016 0.024 0.04
成分 配制不同体积SDS-PAGE分离胶所需各成分的体积(毫升)
8%胶 5 10 15 20 30 50
蒸馏水 1.7 3.3 5 6.7 10.0 16.7
30%Acr-Bis(29:1) 1.3 2.7 4.0 5.3 8.0 13.3
1M Tris, pH8.8 1.9 3.8 5.7 7.6 11.4 19.0
10%SDS 0.05 0.1 0.15 0.2 0.3 0.5
10%凝胶聚合催化剂 0.05 0.1 0.15 0.2 0.3 0.5
TEMED 0.003 0.006 0.009 0.012 0.018 0.03
成分 配制不同体积SDS-PAGE分离胶所需各成分的体积(毫升)
10%胶 5 10 15 20 30 50
蒸馏水 1.3 2.7 4.0 5.3 8.0 13.3
30%Acr-Bis(29:1) 1.7 3.3 5.0 6.7 10.0 16.7
1M Tris, pH8.8 1.9 3.8 5.7 7.6 11.4 19.0
10%SDS 0.05 0.1 0.15 0.2 0.3 0.5
10%凝胶聚合催化剂 0.05 0.1 0.15 0.2 0.3 0.5
TEMED 0.002 0.004 0.006 0.008 0.012 0.02
成分 配制不同体积SDS-PAGE分离胶所需各成分的体积(毫升)
12%胶 5 10 15 20 30 50
蒸馏水 1.0 2.0 3.0 4.0 6.0 10.0
30%Acr-Bis(29:1) 2.0 4.0 6.0 8.0 12.0 20.0
1M Tris, pH8.8 1.9 3.8 5.7 7.6 11.4 19.0
10%SDS 0.05 0.1 0.15 0.2 0.3 0.5
10%凝胶聚合催化剂 0.05 0.1 0.15 0.2 0.3 0.5
TEMED 0.002 0.004 0.006 0.008 0.012 0.02
成分 配制不同体积SDS-PAGE分离胶所需各成分的体积(毫升)
15%胶 5 10 15 20 30 50
蒸馏水 0.5 1.0 1.5 2.0 3.0 5.0
30%Acr-Bis(29:1) 2.5 5.0 7.5 10.0 15.0 25.0
1M Tris, pH8.8 1.9 3.8 5.7 7.6 11.4 19.0
10%SDS 0.05 0.1 0.15 0.2 0.3 0.5
10%凝胶聚合催化剂 0.05 0.1 0.15 0.2 0.3 0.5
TEMED 0.002 0.004 0.006 0.008 0.012 0.02

注:如果配制非变性胶,参考上述配方,不加10%SDS即可配制成非变性PAGE胶。
3.按照如下表格配制SDS-PAGE的浓缩胶(也称堆积胶、积层胶或上层胶):

成分 配制不同体积SDS-PAGE浓缩胶所需各成分的体积(毫升)
5%胶 2 3 4 6 8 10
蒸馏水 1.4 2.1 2.7 4.1 5.5 6.8
30%Acr-Bis(29:1) 0.33 0.5 0.67 1.0 1.3 1.7
1M Tris, pH6.8 0.25 0.38 0.5 0.75 1.0 1.25
10%SDS 0.02 0.03 0.04 0.06 0.08 0.1
10%凝胶聚合催化剂 0.02 0.03 0.04 0.06 0.08 0.1
TEMED 0.002 0.003 0.004 0.006 0.008 0.01

注:通常10-30分钟内胶会凝固。注:具体的凝固时间和温度及光照有关,说明书中10%凝胶聚合催化剂和TEMED的正常推荐用量是室温为25℃时的推荐用量。为达到与25℃时相近的凝固时间,当室温低于25℃时,可以适当同时加大10%凝胶聚合催化剂和TEMED的用量,例如20℃时建议用量是正常推荐用量的1.5倍,15℃时建议用量是正常推荐用量的2倍。



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