ChIP-qPCR Analysis of ER-Ligand Interactions

ChIP was performed as described [66 (link)] from MCF-7 cells treated with ERα ligands for various times with the following antibodies: ERα HC-20 (Santa Cruz Biotechnology sc-543 (Dallas, TX, USA)), rabbit IgG (Cedarlane 011-000-003 (Burlington, ON, Canada)), SUMO2/3 (Cedarlane M114-3), or mouse IgG (Cedarlane 015-000-003). The abundance of immunoprecipitated DNA fragments was quantified by qPCR (Light Cycler 480, Roche) with UPL assays (Roche) (Suppl. Table 5). Results were analyzed by the Percent Input Method.

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Traboulsi T., El Ezzy M., Dumeaux V., Audemard E, & Mader S. (2018). Role of SUMOylation in differential ERα transcriptional repression by tamoxifen and fulvestrant in breast cancer cells. Oncogene, 38(7), 1019-1037.

Publication 2018

ERα HC-20 Light Cycler 480 UPL assays

Antibodies Assays Chip Ligands Light Mcf 7 cells Mouse Rabbit

Corresponding Organization :

Other organizations : Institute for Research in Immunology and Cancer, Université de Montréal

Top 5 similar protocols

Variable analysis

independent variables

ERα ligands
Treatment duration

dependent variables

Abundance of immunoprecipitated DNA fragments

control variables

MCF-7 cells

positive controls

ERα HC-20 (Santa Cruz Biotechnology sc-543 (Dallas, TX, USA))
SUMO2/3 (Cedarlane M114-3)

negative controls

Rabbit IgG (Cedarlane 011-000-003 (Burlington, ON, Canada))
Mouse IgG (Cedarlane 015-000-003)

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