目录号:PE001-01A
产品描述:
肠激酶(Enterokinase, light chain)是一种高度专一性识别Asp-Asp-Asp-Asp-Lys序列的蛋白酶,在Lys的C端水解多肽。它可以将胰蛋白酶原转变为胰酶,也可以将带有这个识别序列的融合蛋白切开。本产品为采用重组酵母分泌表达系统制备的高纯度、高活性的牛肠激酶,适用范围广(4-45℃,pH 4.5-9.5),并且在各种去垢剂和变性剂存在的条件下仍具有部分活性。
产品特点:
高纯度:经多次柱纯化,SDS-PAGE检测图谱无杂蛋白条带;
高活性:在各种去垢剂和变性剂存在的条件下仍具有部分活性;
高特异性:不含其它蛋白酶活性,无非特异性酶切。
使用建议:
建议酶切反应缓冲液为20mM Tris-HCl,50mMNaCl,pH 7.5,温度为20℃~25℃。
注意:上述反应条件只是一般建议,不一定适合所有的蛋白,建议客户针对不同的蛋白提前进行预实验,以确定最佳反应条件。
保存温度:
-20℃
注意事项:
1)在>2M Urea,>250mM NaCl,>20mM β-ME,>0.1% SDS,>50mM imidazole条件下,会影响酶切效果。倘若样品溶液中含有上述成分的一种或多种,为获得理想的酶切结果,建议先将样品透析到1×反应缓冲液中,再进行酶切。
2)建议客户针对不同的蛋白提前进行预实验,以确定最佳反应条件。
应用实例:
图例)重组牛肠激酶酶切反应:每个反应添加15μg的Thioredoxin-NP-27和一定量的肠激酶,37℃经过16小时的反应后用15%的SDS-PAGE胶检测酶切效果。
泳道 A:No rEK ;泳道 B:0.0008U;
泳道 C:0.0015 U;泳道 D:0.003U;
泳道 E:0.008 U;泳道 F:0.015U;
泳道 G:0.03 U
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For research use only.