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2 x 100 µg of MCF-7 nuclear extract (H2O2 treated) at 2.5 µg/µl.
Background
MCF-7 nuclear extract (H2O2 treated) was prepared from cell cultures of the human MCF-7 breast adenocarcinoma cancer cell line. The MCF-7 cell line was originally derived from a pleural effusion of a 69-year old Caucasian woman with metastatic breast cancer. MCF-7 cells are the best characterized and most commonly utilized cell lines in in vitro breast cancer studies. The MCF-7 cell line has retained several characteristics that are particular to mammary epithelium, including the ability to grow as a monolayer, form domes, and respond to hormones. These cells also display a low metastatic potential, leading to the assumption that they represent an early epithelial adenocarcinoma phenotype. These characteristics make MCF-7 cells an ideal model system to study malignant progression in relation to breast cancer.
Treatment of MCF-7 cells with reactive oxygen species (ROS), such as H2O2 triggers an oxidative stress response that leads to toxicity and eventual apoptosis. H2O2 signals an antiproliferative/repair response that induces antioxidant systems to metabolize H2O2 to avert ROS toxicity.
Application Notes
MCF-7 nuclear extract (H2O2 treated) is recommended for studies related to 1) breast cancer research, 2) cellular response to oxidative stress and 3) apoptosis.
Extract Origin
Human breast carcinoma
Extract Composition
MCF-7 nuclear extract in collected in Lysis Buffer after a 3-hr incubation with 0.2 mM H2O2. The Lysis Buffer consists of 20 mM Hepes (pH 7.9), 100 mM KCl, 1 mM MgCl2, 20% glycerol, 0.5 mM PMSF and 0.5 mM DTT. The protein content has been determined by a Bradford-based assay.
Quality Control
Each lot has been tested for p53 activation by using TransAM® p53 Kit. The signal intensity for p53 activation in each lot is compared to the signal intensity obtained with extracts from untreated MCF-7 cells (see figure). Once the signals are blanked, the ratio between the signals from extracts of treated cells over untreated cells used at 5 µg/well must be above 4. This ratio may vary depending of basal level of p53 activation in a given cell type.
Figure 1: Monitoring p53 activation with the TransAM p53 Kit.
Different amounts of nuclear extracts from untreated (opened squares) and H2O2-treated (filled circles) MCF-7 cells are tested for p53 activation using the TransAM p53 Kit.
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.
