GST Pulldown Assay for ERα Interaction

The GST pulldown assay was carried out as previously described^{58 (link)}. The GST alone and GST-fusion proteins were expressed in Escherichia coli BL21 (Takara) and purified using Pierce GST Spin Purification Kit (Thermo scientific) as per the manufacturer’s instruction. The purified proteins were subjected to immobilized on the Pierce Spin Column and then preyed ERα from MCF-7 cells lysate.

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Xu Z., Yang Y., Li B., Li Y., Xia K., Yang Y., Li X., Wang M., Li S, & Wu H. (2018). Checkpoint suppressor 1 suppresses transcriptional activity of ERα and breast cancer cell proliferation via deacetylase SIRT1. Cell Death & Disease, 9(5), 559.

Publication 2018

Escherichia coli BL21 Pierce GST Spin Purification Kit

Assay Escherichia coli Mcf 7 cells Proteins

Corresponding Organization :

Other organizations : Dalian University, Dalian University of Technology

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Variable analysis

independent variables

Expression of GST alone and GST-fusion proteins in Escherichia coli BL21 (Takara)

dependent variables

Binding of purified GST alone and GST-fusion proteins to immobilized ERα from MCF-7 cells lysate

control variables

Purification of GST alone and GST-fusion proteins using Pierce GST Spin Purification Kit as per the manufacturer's instruction
Immobilization of purified proteins on the Pierce Spin Column

controls

Positive control: GST-fusion proteins binding to ERα from MCF-7 cells lysate
Negative control: GST alone binding to ERα from MCF-7 cells lysate

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