Detection and Manipulation of the Stress Response Protein Metallothionein.
Metallothioneins (MTs) are small molecular weight stress response proteins that play a central role as reservoir of essential divalent heavy metal cations such as zinc and copper, and also can diminish the effects of toxic heavy metals such as mercury and cadmium. Historically, MT has been considered to be an intracellular protein with roles to play in the management of heavy metals, as a regulator of cellular redox potential, and as a buffer of free radicals. Our recent studies have highlighted immunomodulatory role of MT in inflammatory diseases and also in the progression of metastatic cell movement. Hence, manipulation and detection of MT is essential for its possible use as a diagnostic and in therapeutic interventions of chronic inflammation. This review describes procedures used to detect MT using techniques such as western immunoblot, competition ELISA, flow cytometry and immunohistochemistry. Additionally, it also describes the use of a colorimetric cell proliferation assay (CellTiter 96 AQueousOne Solution/MTS) to study the proliferative effect of MT. © 2017 by John Wiley & Sons, Inc.
Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut.
Sadikshya
S
Bhandari
Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut.
Clare
C
Melchiorre
Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut.
Kristen
K
Dostie
Department of Internal Medicine, De Pintelaan, Gent, Belgium.
Debby
D
Laukens
Department of Internal Medicine, De Pintelaan, Gent, Belgium.
Lindsey
L
Devisscher
StressMarq Biosciences, Victoria, British Columbia, Canada.
Ariel
A
Louwrier
Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut.
Amy
A
Thees
Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut.
Michael A
MA
Lynes
D000818:Animals; D015153:Blotting, Western; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D008668:Metallothionein / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D018384:Oxidative Stress; D016612:Paraffin Embedding; D011487:Protein Conformation; D013154:Spleen / Q000166:cytology
D008668:Metallothionein
inflammation; metal homeostasis; metallothionein; stress