Jurkat nuclear extract (TPA + CI stimulated)


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Cat.No.:  EL-0381
Product Name:  Jurkat nuclear extract (TPA + CI stimulated)
Description:  Jurkat nuclear extract (TPA + CI stimulated) was prepared from cell cultures of the Jurkat human CD4(+) T cell lymphoblast-like cell line. The Jurkat cell line was established from the propagation of peripheral blood cells of a 14 year old boy with T cell leukemia. These cells are primarily used in research as a T cell model system, particularly in studies of T cell signaling, T cell leukemia, and response to viral infection. Jurkat cells are frequently used in HIV-related research to examine the mechanisms governing T cell-mediated immunity in response to HIV infection. Stimulation of these cells with phorbol esters, such as 12-O-tetradecanoylphorbol-13-acetate (TPA) and either lectins, such as phytohaemagglutinin (PHA) or monoclonal antibodies leads to activation of T cell antigen receptors and induction of IL-2 expression. Because these cells can produce IL-2 in response to stimulation, they are frequently used in research to study the regulation of cytokine expression and differentiation. However, the primary use of Jurkat cells is to study the response of various cancers to radiation or drug treatments. Double stimulation of Jurkat cells with calcium ionophore (CI) the phorbol ester TPA activates T cell antigen receptor (TCR)-mediated signaling and induces the production of large amounts of IL-2. This double stimulation also activates at least two downstream signaling pathways that leads to an increase in intracellular calcium and upregulation of phosphorylation by serine and tyrosine protein kinases and protein kinase C. Increased phosphorylation events lead to modifications in intracellular activity, including changes in gene transcription, such as the induction of expression of IL-2R P55, and activation of regulatory proteins, such as phospholipase-Cγ1 (PLCγ1), a regulator of calcium influx, and NFATc1.
Species:  Human
Source:  Human T lymphoblast; acute T cell leukemia
Components:  2 x 100 µg of Jurkat nuclear extract (TPA + CI stimulated) at 2.5 µg/µl.
Extract Composition:  Jurkat nuclear extract (TPA + CI stimulated) is supplied in Dilution Buffer (20 mM Hepes (pH 7.9), 100 mM KCl, 1 mM MgCl2, 20% glycerol, 0.5 mM PMSF and 0.5 mM DTT). Cells were cultured in medium supplemented with 50 ng/ml TPA (phorbol, 12-myristate, 13 acetate) and 0.5 µM calcium ionophore A23187 (CI) for two hours at 37°C immediately prior to harvesting.
Application Notes:  Jurkat nuclear extract (TPA + CI stimulated) is specifically recommended for use in studies related to 1) TCR-mediated signaling, 2) T cell-mediated immunity and response to viral infection (e.g. HIV), 3) cytokine induction and cell differentiation, and 4) cellular response to cancer therapies.
Storage:  Store at -80°C. Aliquot to avoid repeated freezing and thawing.
Warning:  For research use only, not for diagnostic or therapeutic use.

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