
Application | Each vial is sufficient for a volume of 100ml solution |
---|---|
Just add to your cell lysis buffer |
The Xpert Phosphatase Inhibitor Cocktail Solution(100X)
is a mixture of several inhibitors to prevent valuable
proteinsfrom dephospholylation. Phosphorylation and
dephosphorylation is a molecular on/off switch that regulates
a number of key biological pathways within the cell,
including signal transduction, cell division, and apoptosis.
The Xpert Phosphatase Inhibitor Cocktail preserves
phosphorylation of proteins in cell lysates and tissue
extracts.The cocktail contains a mixture of four
phosphatase inhibitors of broad specificity, including
sodium fluoride, sodium orthovanadate, sodium
pyrophosphateand sodium glycerophosphate.
Unlike other commerciallyavailable phosphatase
inhibitor cocktails which protectagainst either
Ser/Thr phosphatases or protein tyrosine
phosphatases.
The Xpert Phosphatase Inhibitor Cocktail protects
phosphoproteins from both families of phosphatases.
Conveniently packaged in a 100X format, the cocktail
is stable at 4°C and is compatible with common protein
assays, including the BCA Protein Assay and Bradford-
basedassays such as Coomassie.
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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.

