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Cell, Acid & Tissue Extracts

Jurkat nuclear extract (TPA + CI stimulated)

Catalog No: 36013 Format: 200 µg $165 Buy Now

for Jurkat nuclear extract (TPA + CI stimulated)Contents

2 x 100 µg of Jurkat nuclear extract (TPA + CI stimulated) at 2.5 µg/µl.

for Jurkat nuclear extract (TPA + CI stimulated)Background

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.

for Jurkat nuclear extract (TPA + CI stimulated)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.

for Jurkat nuclear extract (TPA + CI stimulated)Extract Origin

Human T lymphoblast; acute T cell leukemia

for Jurkat nuclear extract (TPA + CI stimulated)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.

for Jurkat nuclear extract (TPA + CI stimulated)Quality Control

Extracts have been quality control tested by Western blot and TransAM® Transcription Factor ELISAs.

 
NFκB family profiling of DNA binding activation in various cell lines.

Figure 1: NFκB family profiling of DNA binding activation in various cell lines.
Nuclear extracts prepared from HeLa, HeLa treated with TNF-α, Jurkat, Jurkat treated with PMA and calcium ionophore, and Raji cells were assayed at 10 µg/well for p65, p50, c-Rel, p52 and RelB activity using the TransAM NFκB Family Kit.

for Jurkat nuclear extract (TPA + CI stimulated)Storage

To ensure stability, extracts should be stored at -80°C.

We recommend aliquoting the extracts into single-use fractions and then storing them at -80°C. This eliminates repeated freeze/thaw cycles.

for Jurkat nuclear extract (TPA + CI stimulated)Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

 

Technical Data Sheet

Jurkat nuclear extract (TPA + CI stimulated)
 

Data Thumbnails

NFκB family profiling of DNA binding activation in various cell lines.

Assay of NFκB family activation using the TransAM NFκB Family Kit.
(Click image to enlarge and see details.)