WI-38 nuclear extract (TPA + CI stimulated)
for WI-38 nuclear extract (TPA + CI stimulated)Contents
2 x 100 µg of WI-38 nuclear extract (TPA + CI stimulated) at 2.5 µg/µl.
for WI-38 nuclear extract (TPA + CI stimulated)Background
WI-38 nuclear extract (TPA + CI stimulated) was prepared from a cell culture of the human lung fibroblast WI-38 cell line. The WI-38 cell line was originally derived from lung tissue obtained from a Caucasian female fetus of 3 months gestation. The WI-38 cell line was the first normal diploid human cell line to be continuously cultivated. WI-38 cells are known to have the broadest viral spectra of any cell population that has been tested and are frequently used in research to isolate viruses, such as rhinovirus, and for the production of viral vaccines, including rubella, rabies, and hepatitis A. These cells have a limited replicative lifespan of approximately 50 population doublings after which they enter a state of irreversible growth arrest known as replicative senescence. WI-38 cells are most commonly used in research in studies of viral production, replicative senescence, and as normal controls.
Treatment of cells with the potent tumor-promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) leads to various biological changes that mimic those observed in cells transformed by chemical carcinogens or viruses. Double stimulation of WI-38 cells with calcium ionophore (CI) the phorbol ester TPA activates 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 leads 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-C?1 (PLC?1), a regulator of calcium influx, and NFATc1.
for WI-38 nuclear extract (TPA + CI stimulated)Application Notes
WI-38 nuclear extract (TPA + CI stimulated) is specifically recommended for analysis of the mechanisms underlying tumorigenesis or TPA-mediated tumor conversion and its associated biochemical responses, including increased membrane metabolism, induction of protein biosynthesis, and alterations in the cell cycle.
for WI-38 nuclear extract (TPA + CI stimulated)Extract Origin
Human normal lung fibroblast
for WI-38 nuclear extract (TPA + CI stimulated)Extract Composition
WI-38 nuclear extract was collected in Lysis Buffer after a 2-hour incubation with TPA (100 ng/ml) and calcium ionophore (2 µg/ml). The Lysis Buffer consists of 20 mM Hepes pH 7.5, 400 mM NaCl, 20% glycerol, 0.1 mM EDTA, 10 mM NaF, 10 µM Na2MoO4, 1 mM NaVO3, 10 mM PNPP, 10 mM ?-glycerophosphate, 1 mM DTT and protease inhibitors. The protein content has been determined by a Bradford-based assay.
for WI-38 nuclear extract (TPA + CI stimulated)Quality Control
Each lot has been tested for AP-1 activation by using TransAM® AP-1 Family Kits. Incubation with wild-type competitor oligos reduces binding by over 80% while incubation with mutant oligos had no effect on AP-1 Family binding to DNA (see figure).
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 WI-38 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.