How does oriscience phosphatase inhibitor cocktail help maintain the phosphorylation state of proteins?
Publish Time: 2025-07-23
In modern life science research, protein phosphorylation modification is considered to be one of the core mechanisms for regulating processes such as cell signaling, metabolic regulation, stress response and disease occurrence. However, during the cell or tissue lysis and extraction process, endogenous phosphatases will quickly catalyze the dephosphorylation of phosphorylated proteins, causing the originally phosphorylated protein signal to weaken or even disappear, seriously affecting the accuracy and repeatability of experimental results. To solve this problem, researchers usually add phosphatase inhibitors during sample processing. oriscience phosphatase inhibitor cocktail is designed for this purpose. It effectively inhibits the activity of various types of phosphatases through dual targeting and broad-spectrum coverage, thereby helping to maintain the phosphorylation state of proteins and ensuring the stability and reliability of experimental data.The necessity of phosphatase inhibitors: Why prevent protein dephosphorylation?In cell extracts or tissue samples, there are many phosphatases such as acid phosphatase (ACP), alkaline phosphatase (ALP), protein tyrosine phosphatase (PTP) and serine/threonine phosphatase (PPs). These enzymes can quickly remove phosphate groups from target proteins, resulting in loss of phosphorylation signals.Especially when performing WB, IP, Co-IP, pull-down, IHC, IF and other experiments, if the phosphorylated protein in the sample is dephosphorylated, it will directly affect the detection sensitivity and specificity of the target protein. Therefore, adding efficient phosphatase inhibitors to the sample lysate is a key step to maintain the phosphorylation state of the protein and improve the quality of the experiment.Composition and function of oriscience phosphatase inhibitor cocktailoriscience phosphatase inhibitor cocktail is divided into two parts, tube A and tube B. Each component has been carefully screened to achieve broad-spectrum inhibition of different phosphatase types.Tube A (100×, dissolved in ddH₂O) contains 5 inhibitors: sodium fluoride, sodium orthovanadate, sodium tartrate dihydrate, sodium molybdate and imidazole. These components mainly target acid phosphatases, alkaline phosphatases and protein tyrosine phosphatases (PTPs), and are suitable for the stabilization of tyrosine phosphorylated proteins in various cell signaling pathways.Tube B (100×, DMSO solution) contains 3 inhibitors: (-)-p-bromotetraimidazole oxalate, cantharidin and calyculin A. They mainly act on serine/threonine phosphatases (such as PP1, PP2A) and alkaline phosphatases, effectively maintaining the phosphorylation state of serine and threonine sites.This A+B combination design enables the Oriscience mixture to comprehensively cover different types of phosphatases, thereby preventing protein dephosphorylation to the greatest extent.How to use oriscience phosphatase inhibitor cocktail?The method of use is simple and efficient: add the solutions of tubes A and B respectively in a ratio of 1:100 to the cell lysate or tissue extract to immediately exert the inhibitory effect. Since tube A is an aqueous solution and tube B is a DMSO solution, it is recommended to mix thoroughly before adding and adjust the final solvent concentration according to the experimental system to avoid interference with downstream experiments.In addition, users can also use tube A (Cat. No. PC203) or tube B (Cat. No. PC204) separately according to experimental requirements to meet the needs of specific research directions or target specific phosphatase types.Applied to a variety of experimental scenarios to improve data reliabilityoriscience phosphatase inhibitor cocktail is widely used in a variety of experimental scenarios such as protein immunoblotting (WB), immunoprecipitation (IP), co-immunoprecipitation (Co-IP), pull-down, immunohistochemistry (IHC), immunofluorescence (IF) and kinase activity research. Whether in basic research or disease mechanism exploration, this product can effectively protect phosphorylated proteins from degradation, thereby helping researchers to more accurately capture signal changes and reveal key regulatory nodes.Phosphorylation modification is a "switch" in the cell regulatory network, and its dynamic changes are of great significance to scientific research. And oriscience phosphatase inhibitor cocktail is a key tool to help researchers "lock" these changes. Through the A+B double-tube design, it achieves efficient inhibition of multiple phosphatases, which not only improves the stability of the experiment, but also enhances the credibility of the data. oriscience phosphatase inhibitor cocktail is a powerful assistant for scientific researchers in their phosphorylation research.
