
Inhibited Protease | Serine, Cysteine, Aspartic, and Metalloprotease |
---|---|
Downstream App | No interfering such as 2-D gel and His-tagged protein purification |
EDTA Free | 0.5M EDTA solution supplied in a seperate vial |
The Xpert Protease Inhibitor Cocktail Solution(100X) is
a specially formulated mixture of 5 protease inhibitors
with broad specificity for the inhibition of various protease
and esterase. Other EDTA-free inhibitor cocktails only
provide protection against serine and cysteine protease.
The Xpert Protease inhibitor cocktail solution have
a proprietary form of pepstatin A and Bestatin that allow
for additional protection against aspartic and
metalloprotease without interfering with downstream
applications, such as 2-D gel and His-tagged protein
purifications. It is provided as either a set of 5,10 vials
or as a single vial with 0.5M EDTA.
The Xpert Protease Inhibitor Cocktail Solution(100X) is
ready-to-use 100X stock solution and does not require
reconstitution with water or solvent.
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最好是测浓度,一般不超过30-40ug,上样缓冲液一般4×或者5×,这个没太大区别。
1。称量氨基丁三醇 242g,Na2EDTA.2H2O 37.2g 于1L烧杯中;
2。向烧杯中加入约600ml去离子水,充分搅拌均匀;
3。加入57.1ml的冰乙酸,充分溶解;
4。用NaOH调pH至8.3,加去离子水定容至1L后,室温保存。
使用时稀释50倍 即1×TAE Buffer
10×TBE Buffer配制方法:
1。称量氨基丁三醇 108g,Na2EDTA.2H2O 7.44g,硼酸55g 于1L烧杯中;
2。加入约700ml去离子水,搅拌均匀;
3。用NaOH调pH至8.3,加去离子水定容至1L后,室温保存。
使用时稀释10倍 即1×TBE Buffer
2乘SDS-PAGE上样缓冲液(20ml体系)
1.0mol/L Tris-HCL (pH 6.8) 2ml
1.0mol/L DTT 4ml
SDS 0.8g
溴芬蓝 0.04g
甘油 4ml
dd水 定容至20ml
4℃冰箱保存,可以不分装,我们实验室一般用三个月以上
先配1.0mol/L的DTT,双蒸无菌水溶解4℃冰箱保存,配上样缓冲液的时候加入就可以了,都混匀保存也没问题的
配制分三步: 1) 1 M Tris-HCl (pH 8.0) 50 ml的配制:称取Tris碱6.06 g,加超纯水40 ml溶解,滴加浓HCl约2.1 ml调pH至8.0,定容至50 ml。
① 上样缓冲液中包括甘油,溴酚蓝,SDS等,甘油主要作用是防止样品漂浮出点样孔,溴酚蓝作为电泳指示剂,用来观察电泳进度,SDS用于一些核酸结合蛋白的变性解离.
②核酸染料与凝胶中核酸结合,以便于在紫外光下显示核酸条带.

