How do you dilute ionomycin?

Preparation of 10 mg/ml (13.38 mM) stock solution Add 100 μl of DMSO (not provided) to 1 mg of ionomycin. Mix by vortexing until completely dissolved. 2. Prepare further dilutions with endotoxin‑free water.

What does ionomycin do to cells?

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 cells [1]. Ionomycin can be used to increase intracellular calcium levels triggering cell death through apoptosis and autophagy [1].

How do you store ionomycin?

Storage/Stability Store the powdered ionomycin product desiccated and protected from light at 2–8 °C. Under these conditions the product is stable for 3 years. Stock solutions in ethanol or DMSO, stored at –20 °C and protected from light, are stable for several months.

What is LPS stimulation?

Lipopolysaccharide (LPS) stimulates immune responses by interacting with the membrane receptor CD14 to induce the generation of cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-1, and IL-6.

How do you dissolve thapsigargin?

For maximum solubility in aqueous buffers, thapsigargin should first be dissolved in ethanol and then diluted with the aqueous buffer of choice. Thapsigargin has a solubility of approximately 0.3 mg/ml in a 1:2 solution of ethanol:PBS (pH 7.2) using this method.

Why are Jurkat cells used?

Jurkat cell model can simulate the function T lymphocytes, so it is widely used in the in vitro studies of T cell signal transduction, cytokines, and receptor expression, and can provide reference and guidance for the treatment of various infectious diseases and the research on their pathogenesis.

What do Jurkat cells express?

Jurkat cells are an immortalized line of human T lymphocyte cells that are used to study acute T cell leukemia, T cell signaling, and the expression of various chemokine receptors susceptible to viral entry, particularly HIV.

What is the function of thapsigargin?

Thapsigargin is an inhibitor of sarco endoplasmic reticulum Ca2 ATPase (SERCA). It pumps calcium ions from the cytoplasm into the lumen of the endoplasmic reticulum (ER) and thapsigargin. This process will causes an increase in the cytoplasmic calcium levels while also depleting ER stores.

How much DMSO is needed to make ionomycin?

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.

How do you make ionomycin from calcium salt?

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.

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.

What is the concentration of ionomycin calcium salt solution i3909?

It is recommended to store I3909 under an inert gas and at -20°C protected from light. What is the concentration of the Ionomycin calcium salt solution, Product No. I3909? It is a 1mM solution or ~ 0.75mg/mL in DMSO.