Immunoblot and Immunofluorescence Assay Protocol

Immunoblot and immunofluorescence assays were performed as previously described [29 (link)]. Antibodies employed to perform these studies were as follows: centrin (20H5 kindly provided by Dr. Salisbury’s laboratory at the Mayo Clinic); ERα and pericentrin (Santa Cruz Biotechnology, Dallas, TX, USA); AURKA (Cell Signaling Technology, Danvers, MA, USA); CD44, CD24, NOTCH1, NOTCH2, and NOTCH3 (Abcam, Cambridge, MA, USA); and β-actin and α-tubulin (Sigma-Aldrich, St. Louis, MO, USA). Secondary antibodies were obtained from Molecular Probes (Eugene, OR, USA).

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Leontovich A.A., Jalalirad M., Salisbury J.L., Mills L., Haddox C., Schroeder M., Tuma A., Guicciardi M.E., Zammataro L., Gambino M.W., Amato A., Di Leonardo A., McCubrey J., Lange C.A., Liu M., Haddad T., Goetz M., Boughey J., Sarkaria J., Wang L., Ingle J.N., Galanis E, & D’Assoro A.B. (2018). NOTCH3 expression is linked to breast cancer seeding and distant metastasis. Breast Cancer Research : BCR, 20, 105.

Publication 2018

centrin AURKA NOTCH2 NOTCH3 β-actin and α-tubulin

Actin Antibodies Aurka Cd44 Centrin Immunoblot Immunofluorescence assays Molecular probes Notch2 Notch3 Pericentrin α tubulin

Corresponding Organization :

Other organizations : Mayo Clinic, Yale University, University of Palermo, East Carolina University, University of Minnesota

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Variable analysis

independent variables

Antibodies employed to perform these studies

dependent variables

Immunoblot and immunofluorescence assays

control variables

Immunoblot and immunofluorescence assays were performed as previously described [29 (link)]
Secondary antibodies were obtained from Molecular Probes (Eugene, OR, USA)

positive controls

Centrin (20H5 kindly provided by Dr. Salisbury's laboratory at the Mayo Clinic)

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