For Phosphorylation Protection, more inhibitors mean better results. So we provide Two Tubes with 8 inhibitors. Phosphorylation of proteins and lipids is a hallmark of the signaling or metabolic status of cells. For decades, researchers have used easily-accessible phosphate mimetics (tube A) to prevent the loss of phosphorylation on biomolecules in vitro. In recent years however, a number of naturally derived agents (tube B) have been discovered as potent inhibitors of phosphatases, thus increasing the available protection spectrum.
Contents
Tube #
Ingredient
100x Conc.
Target
A (Aqueous)
Sodium Fluoride
100 mM
Acid phosphatases
Sodium Orthovanadate
100 mM
Alkaline phosphatases, PTPs, ATPases
Sodium Tartrate
400 mM
Acid phosphatases
Sodium Molybdate
115 mM
Acid and phosphoprotein phosphatases
Imidazole
200 mM
Alkaline phosphatases
B (DMSO)
(-)-p-Bromotetramisole oxalate
2.5 mM
Alkaline phosphatases
Cantharidin
500 μM
Ser/Thr phosphatases
Microcystin LR, Microcystis aeruginosa
500 nM
PP1 and PP2A
Works well in common detergents such as 1% SDS, Triton, and NP-40 DTT, EDTA and EGTA may reduce the protection effects of Sodium Orthovanadate
储存条件
The product should be stored at -20°C, and is stable for up to 12 months. When kept at 2-8°C, the product remains stable for 2 months.
实验方法
Application Fields
Western Blot analysis, Co-IP, pull-down, IF, IHC, kinase assay and etc.
Experiment Protocol
Thaw at room temperature. Before assaying, add each solution at 1:100 (v/v) dilution to samples (such as cell lysates or tissue extracts). Mix the sample sufficiently after each time of addition.
Note: For optimum protection of phosphorylated proteins and lipids, we recommend simultaneous use of BOTH Tubes A and B. Because the inhibitor cocktails target different families of phosphatases, synergistic stabilization effects are observed with dual-treatment.
The Phosphatase Inhibitor Cocktail does not contain protease inhibitors. Therefore,we recommend that a protease inhibitor cocktail is added beforehand. Protease inhibitor cocktails are frequently used in mammalian cell lysates or tissue extracts to increase protein stability. As the name indicates, the cocktail functions to inhibit proteases that would degrade either phosphorylated or non-phosphorylated protein substrates.