NIH/3T3 whole-cell extract (TPA stimulated)

Catalog No: 36415 Format: 200 µg ¥44,000 Buy

Contents

200 µg of NIH/3T3 whole-cell extract (TPA stimulated) at 5.0 µg/µl.

Background

NIH/3T3 whole-cell extract (TPA stimulated) was prepared from a cell culture of the NIH/3T3 mouse fibroblast cell line obtained from Swiss mouse embryo tissue. This cell line was established from immortalized primary mouse embryonic fibroblast cells that were cultured by the '3T3 protocol'. 3T3 (3-day transfer, inoculum 3 x 105 cells) refers to the transfer and inoculation protocol used to initiate and propagate the cell line. In research, these cells have become the standard mouse fibroblast cell line.

Treatment of NIH/3T3 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. In mice, TPA has been shown to induce long lasting effects, including inflammation and hyperplasia, that are associated with the critical first step in multistage tumorigenesis known as tumor conversion. Thus, TPA stimulated NIH/3T3 cells are frequently utilized in cancer research as a model system for studying the radiomimetic and tumor-promoting activity of TPA on replicating cells, particularly in relation to PKC-mediated growth control.

Application Notes

NIH/3T3 whole-cell extract (TPA 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.

Extract Origin

Mouse embryo

Extract Composition

NIH/3T3 whole-cell extract (TPA 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) for 2 hours at 37°C immediately prior to harvesting.

Quality Control

Extracts have been quality control tested by Western blot and the Electrophoretic Mobility Shift Assay (EMSA).

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.

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.