Why does PMA stimulate ionomycin?
PMA activates protein kinase C, while ionomycin is a calcium ionophore, and stimulation with these compounds bypasses the T cell membrane receptor complex and will lead to activation of several intracellular signaling pathways, resulting in T cell activation and production of a variety of cytokines.
How do you dissolve ionomycin?
1. Add 100 μl of DMSO (not provided) to 1 mg of ionomycin. Mix by vortexing until completely dissolved.
How do you make a PMA solution?
Preparation of stock solution (5 mg/ml) – Add 1 ml of DMSO and vortex until completely dissolved. – Prepare serial dilutions using endotoxin-free water. Note: PMA solutions may remain cloudy. PMA-induced activation of NF-κB PMA can be used as a positive control with NF-κB reporter cell lines.
What is PMA function?
Phorbol myristate acetate (PMA; phorbol ester) activates protein kinase C, which then activates a wide range of signaling pathways, including ERK via effects on the upstream kinase Raf. ERK then activates a wide range of downstream targets, including S6 ribosomal protein.
How do you take ionomycin?
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.
What is PMA diluted in?
4 To prepare aqueous solutions of PMA, it is best to start out with a concentrated solution of the compound in DMSO (20 mM) and dilute a very small aliquot of this solution rapidly with water or buffer.
How do PMA and ionomycin induce glioblastoma cell death?
PMA and ionomycin induce glioblastoma cell death: activation-induced cell-death-like phenomena occur in glioma cells. Phorbol myristate acetate (PMA) and ionomycin (Io) can induce T cell activation and proliferation. Furthermore, they stimulate activation-induced cell death (AICD) in mature lymphocytes via Fas/Fas ligand (FasL) up-regulation.
How do phorbol myristate acetate and ionomycin activate T cells?
Phorbol myristate acetate (PMA) and ionomycin (Io) can induce T cell activation and proliferation. Furthermore, they stimulate activation-induced cell death (AICD) in mature lymphocytes via Fas/Fas ligand (FasL) up-regulation.
What is the difference between Pha and PMA/Iono?
PHA and PMA/Iono act on cells in a quite different manner. PHA (Phytohaemaglutinin) is a lectin, binds to the sugars on glycosylated surface proteins, including the TCR, and thereby crosslinks them.
What is the function of ionomycin?
Ionomycin is a membrane permeable calcium ionophore produced by the bacterium Streptomyces conglobatus. This ionophore facilitates the transfer of calcium ions (Ca2+) into and out of cells1. Ionomycin can be used to increase intracellular calcium levels triggering cell death through apoptosis and autophagy1.