MCF-7 nuclear extract (H2O2 post-treated)

Synonyms

MCF-7, MCF7, MCF 7, MCF-7 (H2O2 treated), H2O2, treated, stimulated, nuclear extract, nuclear, extract, lysate, nuclear lysate, breast cancer, adenocarcinoma, mammary, estrogen, estrogen receptors, ER, breast cancer research, epithelial adenocarcinoma, tumor, cancer, malignant, malignancy, toxicity, ROS, reactive oxygen species, oxidative stress, hydrogen peroxide, antioxidant, antioxidants, apoptosis, programmed cell death, cell death, h202 metabolism, antiproliferative/repair response, cell death, catalase, 40810

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Contents

2 x 100 µg of MCF-7 nuclear extract (H₂O₂ post treated) at 2.5 µg/µl.

Background

MCF-7 nuclear extract (H₂O₂ post-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 post-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 was collected in Lysis Buffer. Post-collection treatment consists of exposing the nuclear extract to a final concentration of 200 mM H₂O₂ for 20 minutes on ice. 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 Sp1 activity by using the TransAM^® Sp1 Kit. The signal intensity for Sp1 activity in each lot is compared to the signal intensity obtained with H₂O₂-post treated extracts from MCF-7 cells (see figure). Once the signals are blanked, the ratio between the signals from untreated extracts over treated extracts used at 5 µg/well must be above 4.

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