S'identifier

Seattle University

3 ARTICLES PUBLISHED IN JoVE

image

Biology

Biochemical Reconstitution of Steroid Receptor•Hsp90 Protein Complexes and Reactivation of Ligand Binding
Patrick J. M. Murphy 1, Hannah R. Franklin 2, Nathan W. Furukawa 3
1College of Nursing, Interdisciplinary Life Sciences Research Laboratory, Seattle University, 2College of Science and Engineering, Interdisciplinary Life Sciences Research Laboratory, Seattle University, 3School of Medicine, University of Washington

An in vitro method for preparing functional glucocorticoid receptor (GR)•hsp90 protein complexes from purified proteins and cellular lysates is described. The method utilizes immunoadsorption of recombinant GR followed by salt-stripping and protein complex reconstitution. The importance of cofactors and buffer conditions are discussed, as are potential method applications.

image

Biology

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Patrick J. M. Murphy 1, Orrin J. Stone 2, Michelle E. Anderson 1
1College of Nursing, Interdisciplinary Life Sciences Research Laboratory, Seattle University, 2College of Science and Engineering, Interdisciplinary Life Sciences Research Laboratory, Seattle University

An automated method for identifying suitable hydrophobic interaction chromatography (HIC) media to be used in the process of protein purification is presented. The method utilizes a medium-pressure liquid chromatography system including automated buffer blending, dynamic sample loop injection, sequential column selection, multi-wavelength analysis, and split fraction eluate collection.

image

Bioengineering

Visualization of Recombinant DNA and Protein Complexes Using Atomic Force Microscopy
Patrick J. M. Murphy 1, Morgan Shannon 2, John Goertz 2
1College of Nursing, Interdisciplinary Life Sciences Research Laboratory, Seattle University, 2College of Science and Engineering, Interdisciplinary Life Sciences Research Laboratory, Seattle University

A tapping mode atomic force microscope (AFM) method for the visualization of plasmid DNA, cytoplasmic proteins, and DNA-protein complexes is described. The method includes alternate approaches for preparing samples for AFM imaging following biochemical manipulation. DNA containing specific protein interacting regions are observed in near-physiologic buffer conditions.

JoVE Logo

Confidentialité

Conditions d'utilisation

Politiques

Recherche

Enseignement

À PROPOS DE JoVE

Copyright © 2024 MyJoVE Corporation. Tous droits réservés.