細胞或組織等提取物中含有許多內源性的蛋白酶、磷酸酶等，容易導致提取物中的蛋白降解或去修飾，從而影響后續(xù)的蛋白檢測。因此在提取物中添加適當的蛋白酶、磷酸酶等抑制劑是防止蛋白降解和去修飾的有效方法。本磷酸酶抑制劑混合物可以有效抑制常見磷酸酶對于蛋白的去磷酸化作用，保持蛋白原有的磷酸化狀態(tài)。
本產品為經優(yōu)化和測試的用于動物細胞或組織、真菌、細菌以及植物蛋白提取的磷酸酶抑制劑混合物，包含了酪氨酸、酸性及堿性磷酸酶抑制劑。
本磷酸酶抑制劑混合物含200mM imidazole, 100mM sodium fluoride, 115mM sodium molybdate, 100mM sodium orthovanadate 和 400mM sodium tartrate dihydrate，配制在超純水中。
本產品使用便捷，通常以1:100的比例把磷酸酶抑制劑混合物加入裂解液中即可用于樣品蛋白的提取，并有效抑制蛋白的去磷酸化。
Application
The phosphatase inhibitor cocktail improves the yields of intact proteins by adding inhibitors to enzymes that modify proteins present in cell extracts. This product has been optimized and tested for tyrosine protein phosphatases, acid and alkaline phosphatases. It has been tested on cell extracts from animal tissues including bovine liver and human placenta, and cytosolic extract of rabbit muscle.
Biochem/physiol Actions
This mixture contains individual components with specific inhibitory properties. Sodium orthovanadate inhibits a number of ATPases, protein tyrosine phosphatases, and other phosphate-transferring enzymes. Sodium molybdate inhibits acid and phosphoprotein phosphatases. Sodium tartrate inhibits acid phosphatases. Imidazole inhibits alkaline phosphatases.
Caution
As supplied, this product is stable for two years at the recommended 2-8°C storage temperature.
Preparation Note
This cocktail is supplied as a clear aqueous solution. It has been sterile filtered through a 0.2 μm membrane in bottles that have been aseptically filled. One mL will inhibit phosphatase activities found in the 100,000 x g supernatant from human placenta, bovine liver, rabbit muscle, A431, or Jurkat cell extracts at a protein concentration of approximately 5 mg/mL.
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