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Western blot analysis of extracts from BAEC cells, untreated or treated with the indicated concentrations of ionomycin for 1 to 2 minutes. The effect of ionomycin treatment on eNOS phosphorylation at Thr495 was detected using Phospho-eNOS (Thr495) Antibody #9574 (upper). eNOS Antibody #9572 was used as a loading control (lower). Ionomycin-induced Ca2+ influx in BAEC cells results in rapid dephosphorylation of eNOS (Thr495).

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Product Usage Information

Ionomycin, calcium salt is supplied as 2.5 mg powder. Store at or below -20ºC. Before use, dissolve powder in 1.1 ml DMSO to make a 3 mM ionomycin stock solution. For working concentrations of 1 μM-3 μM, dilute DMSO stock 1:3000 to 1:1000. Treat cells with the desired concentration for intended time. Ionomycin is more stable as a powder than in solution; it is therefore recommended to dissolve as close as possible prior to use. Store solution at or below -20ºC.


Storage: Store lyophilized or in solution at -20°C. In lyophilized form, the chemical is stable for 24 months. Once in solution, use within 3 months to prevent loss of potency.

Product Description

Molecular Weight:

747.1 g/mol


Purity:

>99%


Molecular Formula:

C41H70O9 • Ca2+


Ionomycin is a potent and selective calcium ionophore agent (1,2). The molecule acts as a motile Ca2+ carrier and enhances Ca2+ influx by direct stimulation of store-regulated cation entry across biological membranes (3). At the micromolar level, ionomycin can activate Ca2+/Calmodulin dependent kinase and phosphatase to stimulate gene expression (4). In human T cells, ionomycin induces hydrolysis of phosphoinositides and activates PKC to mediate T cell activation (5). Ionomycin treatment of human B cells induces the activation of calcium-dependent endonuclease and results in apoptosis (6). Ionomycin treatment of bovine aortic endothelial cells (BAECs) induces rapid dephosphorylation of eNOS at Thr495 and mediates eNOS activation (7).


1.  Liu, C. and Hermann, T.E. (1978) J Biol Chem 253, 5892-4.

2.  Kauffman, R.F. et al. (1980) J Biol Chem 255, 2735-9.

3.  Morgan, A.J. and Jacob, R. (1994) Biochem J 300 ( Pt 3), 665-72.

4.  Lobo, F.M. et al. (1999) J Immunol 162, 2057-63.

5.  Chatila, T. et al. (1989) J Immunol 143, 1283-9.

6.  Aagaard-Tillery, K.M. and Jelinek, D.F. (1995) J Immunol 155, 3297-307.

7.  Lin, M.I. et al. (2003) J Biol Chem 278, 44719-26.



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