Phosphorylation and dephosphorylation of proteins within cells serve as critical regulatory switches for many important biological processes, including signal transduction, cell proliferation, cell differentiation, and apoptosis. Therefore, controlling phosphatase activity is of significant physiological importance in protein research. Tissues and cells contain abundant zymogens, which are released into the extracellular environment upon lysis and become fully activated, leading to the degradation of surrounding proteins. Consequently, preserving the phosphorylated state of proteins is extremely crucial when studying protein phosphorylation pathways and signaling cascades. This product is a specialized mixture of multiple phosphatase inhibitors, providing broad-spectrum inhibition against non-specific acidic/alkaline phosphatases as well as specific phosphatases such as protein tyrosine phosphatases and serine/threonine protein phosphatases (PPA, PP2A). The product is provided in two vials, Tube A and Tube B. The two solutions target different types of phosphatases, enabling synergistic inhibition. Tube A contains Cantharidin, Bromotetramisole, and Microcystin-LR, provided as ready-to-use solutions in DMSO. It inhibits the L-isoenzyme of alkaline phosphatase and serine/threonine protein phosphatases, such as protein phosphatase 1 and 2A (PP1 and PP2A). Tube B contains Sodium orthovanadate, Sodium molybdate, Sodium tartrate, Imidazole, and Sodium Fluoride, provided as ready-to-use solutions in ddH₂O. It inhibits tyrosine protein phosphatases, acid phosphatases, and alkaline phosphatases. This product is suitable for various applications including Western Blotting (WB), Co-Immunoprecipitation (Co-IP), Pull-down assays, Immunofluorescence (IF), Immunohistochemistry (IHC), and kinase studies. Specifications Cat.No. 20109ES05 / 20109ES20 / 20109ES70 Size 2 mL / 10×2 mL / 100×2 mL Components Components No. Name 20109ES05 20109ES20 20109ES70 20109-A InStab™ Phosphatase Inhibitor Cocktail-A Solution（100×） 1 mL 10×1 mL 100×1 mL 20109-B InStab™ Phosphatase Inhibitor Cocktail-B Solution（100×） 1 mL 10×1 mL 100×1 mL Storage This product should be stored at -25~-15℃ for 1 year or stored at 2~8℃ for 2 months. Notes 1. It is recommended to aliquot the 100× storage solution for freezing to avoid repeated freeze-thaw cycles. Working solutions should be freshly prepared before use. 2. This product does not contain protease inhibitors. Depending on experimental requirements, appropriate protease inhibitors can be added to the samples to be analyzed beforehand. 3. Solution A is dissolved in DMSO; handle it with particular care. 4. Substances such as DTT, EDTA, and EGTA may interfere with Sodium orthovanadate contained in this product. Handle these substances carefully. 5. This product is for research use only. 6. For your safety and health, please wear a lab coat and disposable gloves when handling this product. Instructions Before use, remove this product from the refrigerator and allow it to warm to room temperature. Add Solution A and Solution B to the sample separately at a 1:100 (v/v) ratio, mixing thoroughly after each addition. [Notes]: 1)Solution A is dissolved in DMSO, while Solution B is dissolved in ddH2O. DMSO generally dissolves more slowly than water; therefore, Solution A may require a longer time to fully dissolve compared to Solution B. If the temperature is low, Solution A can be placed in a 25°C water bath to aid dissolution. 2)Mixing Solutions A and B directly before use may cause precipitation. This mixing method should be avoided. It is recommended to add the appropriate amounts of Solution A and Solution B separately to the sample. 3)This product is compatible with up to 1% SDS, Triton X-100, and NP-40. Documents: Safety Data Sheet 20109-A-MSDS-CN20250110 20109-B-MSDS-CN20250110 Manuals 20109_Manual_Ver.EN20250815 Citations & References: [1] Zhu R, Yan T, Feng Y, et al. Mesenchymal stem cell treatment improves outcome of COVID-19 patients via multiple immunomodulatory mechanisms. Cell Res. 2021;31(12):1244-1262. doi:10.1038/s41422-021-00573-y(IF:25.617) [2] Liu H, Zhou Y, Qiu H, et al. Rab26 suppresses migration and invasion of breast cancer cells through mediating autophagic degradation of phosphorylated Src. Cell Death Dis. 2021;12(4):284. Published 2021 Mar 17. doi:10.1038/s41419-021-03561-7(IF:8.469) [3] Chen Z, Xie J, Li Q, et al. Human CYP enzyme-activated clastogenicity of 2-ethylhexyl diphenyl phosphate (a flame retardant) in mammalian cells. Environ Pollut. 2021;285:117527. doi:10.1016/j.envpol.2021.117527(IF:8.071) [4] Sun J, Guo Y, Fan Y, Wang Q, Zhang Q, Lai D. Decreased expression of IDH1 by chronic unpredictable stress suppresses proliferation and accelerates senescence of granulosa cells through ROS activated MAPK signaling pathways. Free Radic Biol Med. 2021;169:122-136. doi:10.1016/j.freeradbiomed.2021.04.016(IF:7.376) [5] Yang BY, Deng GY, Zhao RZ, et al. Porous Se@SiO2 nanosphere-coated catheter accelerates prostatic urethra wound healing by modulating macrophage polarization through reactive oxygen species-NF-κB pathway inhibition. Acta Biomater. 2019;88:392-405. doi:10.1016/j.actbio.2019.02.006(IF:6.638) [6] Zhang Y, Li G, Zhang X, Lin L. ROS-scavenging glyco-nanoplatform for synergistic antibacterial and wound-healing therapy of bacterial keratitis. J Mater Chem B. 2022;10(24):4575-4587. Published 2022 Jun 22. doi:10.1039/d2tb00667g(IF:6.331) [7] Deng L, Li H, Su X, et al. Chlorzoxazone, a small molecule drug, augments immunosuppressive capacity of mesenchymal stem cells via modulation of FOXO3 phosphorylation. Cell Death Dis. 2020;11(3):158. Published 2020 Mar 2. doi:10.1038/s41419-020-2357-8(IF:6.304) [8] Zhang X, Cai Z, Wu M, Huangfu X, Li J, Liu X. Adipose Stem Cell-Derived Exosomes Recover Impaired Matrix Metabolism of Torn Human Rotator Cuff Tendons by Maintaining Tissue Homeostasis. Am J Sports Med. 2021;49(4):899-908. doi:10.1177/0363546521992469(IF:6.203) [9] Gao C, Wang SW, Lu JC, et al. KSR2-14-3-3ζ complex serves as a biomarker and potential therapeutic target in sorafenib-resistant hepatocellular carcinoma. Biomark Res. 2022;10(1):25. Published 2022 Apr 25. doi:10.1186/s40364-022-00361-9(IF:6.148) [10] Chen L, Xu W, Yang Q, et al. Imperatorin alleviates cancer cachexia and prevents muscle wasting via directly inhibiting STAT3. Pharmacol Res. 2020;158:104871. doi:10.1016/j.phrs.2020.104871(IF:5.893)
