Expression and Purification of FAK-FERM and HER2-ICD Proteins

His-tagged avian FAK-FERM (residues 31–405) in modified pET vector (provided by Dr. Michael Eck) was expressed in BL21 (DE3) E. coli (Thermo Fisher) and purified on Ni-NTA resin (Thermo-Fisher) similarly as described (T. Marlowe et al., 2019 (link)). GST-FAK-NT (FERM) proteins in pGEX-4T1 vector was expressed in BL21 (DE3) E. coli and purified on Glutathione Sepharose 4B resin (GE Healthcare) as previously described (Golubovskaya, Finch, & Cance, 2005 (link)). Recombinant HER2-ICD (Intracellular Domain) was purchased from Thermo Fisher (cat#PV3366).

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Stahl E., Nott R., Koessel K., Cance W, & Marlowe T. (2020). Computational-based discovery of FAK FERM domain chemical probes that inhibit HER2-FAK cancer signaling. Chemical biology & drug design, 95(6), 584-599.

Publication 2020

BL21 (DE3) E. coli Ni-NTA resin Glutathione Sepharose 4B resin

Avian E coli Ferm Finch Glutathione Her2 Intracellular Proteins Resin Sepharose 4b Vector

Corresponding Organization : University of Arizona

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Variable analysis

independent variables

Expression of His-tagged avian FAK-FERM (residues 31–405) in modified pET vector in BL21 (DE3) E. coli
Expression of GST-FAK-NT (FERM) proteins in pGEX-4T1 vector in BL21 (DE3) E. coli

dependent variables

Not explicitly mentioned

control variables

Purification of His-tagged avian FAK-FERM (residues 31–405) on Ni-NTA resin
Purification of GST-FAK-NT (FERM) proteins on Glutathione Sepharose 4B resin
Recombinant HER2-ICD (Intracellular Domain) purchased from Thermo Fisher

controls

Positive control: Not specified
Negative control: Not specified

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