Sodium Orthovanadate (Part ACTVO-4) is provided as a potent phosphatase inhibitor that has been pre-activated and depolymerized by pH10 adjustment and boiling cycles until the solution maintains a colorless appearance at equilibrium at pH10§. This preparation depolymerizes sodium orthovanadate and enhances its phosphotyrosyl phosphatase inhibitor potency.§
Activated sodium orthovanadate solution is ready-to-use and can be dispensed directly into cell lysates, or cell culture medium (after filter sterilization if applied over an extended duration; i.e. overnight. Sterilization for cell culture applications is not required for short duration exposure; i.e < 5 hrs).
Pre-activated sodium orthovanadate solution is stable for at least one year when stored at -20oC as provided. Certain buffer components, such as EDTA and oxidants, interact with inhibition potency. HEPES is a preferred buffer for cell lysis solutions to maintain vanadate potency.2
Reference on mechanism of inhibition and assay buffer interactions:
2. Huyer, G., et al: J. Biol. Chem. (1997) 272, 843-851.
Sodium Orthovanadate (Vanadate) Applications:
Maintains tyrosyl phosphorylation in cell lysates and live cell culture (syringe filter sterilization is required for cell culture).
Alkaline phosphatase inhibitor
ATPase inhibitor, including ABC transporters
Use as a stock solution for pervanadate generation
Directions (Phosphatase Inhibition)
Concentration: Recommended experimental sodium orthovanadate (vanadate) concentration for phosphatase inhibition is 1 mM.
Cell lysates: Dispense the 200 mM stock sodium orthovanadate solution directly into cell lysates to make a final concentration of 1 mM.
Cell culture: Syringe sterilize with a 2 micron filter, dispense the 200 mM stock solution to cell culture medium to make a final 1 mM concentration.
Images and Data Tab: Sodium Orthovanadate (Vanadate) complexed with alkaline phosphatase. Ref: (1999) J.Biol.Chem.274: 8351-8354. Source: PDB
Product Citations
Song, L., Zhao, S., Frotscher, M. et. al 2022. Phosphorylation of Focal Adhesion Kinase at Y925: Role in Glia-Dependent and Independent Migration through Regulating Cofilin and N-Cadherin. Mol Neurobiol 59, 3467–3484 (2022). Link to article
Munabi, N.C., England, R.W., Edwards, A.K., Kitajewski, A.A., Tan, Q.K., Weinstein, A., Kung, J.E., Wilcox, M., Kitajewski, J.K., Shawber, C.J. and Wu, J.K., 2016. Propranolol Targets Hemangioma Stem Cells via cAMP and Mitogen-Activated Protein Kinase Regulation. Stem cells translational medicine, 5(1), pp.45-55. Link to article
Bunda, Severa, et al. "Src promotes GTPase activity of Ras via tyrosine 32 phosphorylation." Proceedings of the National Academy of Sciences 111.36 (2014): E3785-E3794. Link to article
Bunda, S., Burrell, K., Heir, P., Zeng, L., Alamsahebpour, A., Kano, Y., ... & Ohh, M. (2015). Inhibition of SHP2-mediated dephosphorylation of Ras suppresses oncogenesis. Nature Communications, 6. Link to article
Selected references on sodium orthovanadate (vanadate) on phosphatase inhibition and applications to cell signaling research.
Feng Y, Bhatt AJ, Fratkin JD, and Rhodes PG. Neuroprotective effects of sodium orthovanadate after hypoxic-ischemic brain injury in neonatal rats Brain Res Bull 76: 102-108, 2008.
Hasegawa Y, Morioka M, Hasegawa S, Matsumoto J, Kawano T, Kai Y, Yano S, Fukunaga K, and Kuratsu J. Therapeutic time window and dose dependence of neuroprotective effects of sodium orthovanadate following transient middle cerebral artery occlusion in rats. J Pharmacol Exp Ther 317: 875-881, 2006.
Hulley PA, Conradie MM, Langeveldt CR, and Hough FS. Glucocorticoid-induced osteoporosis in the rat is prevented by the tyrosine phosphatase inhibitor, sodium orthovanadate. Bone 31: 220-229, 2002.
Kafl HE and Elkashef HA. Effect of sodium orthovanadate on the urinary bladder rings isolated from normal and hyperglycemic rats. Pak J Pharm Sci 19: 195-201, 2006.
Klein A, Holko P, Ligeza J, and Kordowiak AM. Sodium orthovanadate affects growth of some human epithelial cancer cells (A549, HTB44, DU145). Folia Biol (Krakow) 56: 115-121, 2008.
Li L, Eisen AZ, Sturman E, and Seltzer JL. Protein tyrosine phosphorylation in signalling pathways leading to the activation of gelatinase A: activation of gelatinase A by treatment with the protein tyrosine phosphatase inhibitor sodium orthovanadate.Biochim Biophys Acta 1405: 110-120, 1998.
Matsumoto J, Morioka M, Hasegawa Y, Kawano T, Yoshinaga Y, Maeda T, Yano S, Kai Y, Fukunaga K, and Kuratsu J. Sodium orthovanadate enhances proliferation of progenitor cells in the adult rat subventricular zone after focal cerebral ischemia. J Pharmacol Exp Ther 318: 982-991, 2006.
Milward AD, Riba R, Patel B, Oberprieler NG, and Naseem KM. Sodium orthovanadate induced tyrosine phosphorylation of platelet nitric oxide synthase negatively regulates enzyme activity. Biochim Biophys Acta 1760: 1411-1417, 2006.
Morita A, Zhu J, Suzuki N, Enomoto A, Matsumoto Y, Tomita M, Suzuki T, Ohtomo K, and Hosoi Y. Sodium orthovanadate suppresses DNA damage-induced caspase activation and apoptosis by inactivating p53. Cell Death Differ 13: 499-511, 2006.
Pelassy C, Breittmayer JP, and Aussel C. Inhibition of phosphatidylserine synthesis during Jurkat T cell activation. The phosphatase inhibitor, sodium ortho-vanadate bypasses the CD3/T cell receptor-induced second messenger signaling pathway. Eur J Biochem 267: 984-992, 2000.
Preet A, Siddiqui MR, Taha A, Badhai J, Hussain ME, Yadava PK, and Baquer NZ. Long-term effect of Trigonella foenum graecum and its combination with sodium orthovanadate in preventing histopathological and biochemical abnormalities in diabetic rat ocular tissues. Mol Cell Biochem 289: 137-147, 2006.
Tran NP, Park JK, Hong SJ, and Lee CG. Proteomics of proteins associated with astaxanthin accumulation in the green algae Haematococcus lacustris under the influence of sodium orthovanadate. Biotechnol Lett 31: 1917-1922, 2009.
Ueki H, Yamasaki Y, Higo K, Motoyashiki T, Kawabata H, and Morita T. Regulation of cyclic AMP phosphodiesterase activity by particulate protein tyrosine kinase and phosphotyrosine phosphatase activities sensitive to sodium orthovanadate. Biol Pharm Bull 18: 214-218, 1995.
Wo YY, Sheu DS, and Lu CH. Sodium vanadate treatment as a shortcut following alkaline phosphatase cleavage.Biotechniques 25: 350-352, 1998.
Wu DN, Pei DS, Wang Q, and Zhang GY. Down-regulation of PTEN by sodium orthovanadate inhibits ASK1 activation via PI3-K/Akt during cerebral ischemia in rat hippocampus. Neurosci Lett 404: 98-102, 2006.
Ordering
In addition to direct purchase from Fivephoton Biochemicals, this product can be purchased on fishersci.com and shipped directly from Fivephoton Biochemicals to the customer. Sodium Orthovanadate is also available from other distributors of Fivephoton Biochemicals.
Pharma/Biotech customers: This product is available in bulk scale for pharmacological screening applications. We can provide customized and newly formulated lots in bulk scale upon request. Contact customersupport@fivephoton.com.
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